Journal of Integrative Agriculture 2017, 16(0): Available online at ScienceDirect

Save this PDF as:
 WORD  PNG  TXT  JPG

Size: px
Start display at page:

Download "Journal of Integrative Agriculture 2017, 16(0): Available online at ScienceDirect"

Transcription

1 Journal of Integrative Agriculture 2017, 16(0): Available online at ScienceDirect RESEARCH ARTICLE Wheat streak mosaic virus: incidence in field crops, potential reservoir within grass species and uptake in winter wheat cultivars Jana Chalupniková 1, 2, Jiban Kumar Kundu 1, Khushwant Singh 1, Pavla Bartaková 1, Eva Beoni 1 1 Division of Crop Protection and Plant Health, Crop Research Institute, Drnovská 507, Prague, Czech Republic 2 Department of Experimental Plant Biology, Faculty of Science, Charles University in Prague, Albertov 6, Prague, Czech Republic Abstract Wheat streak mosaic virus (WSMV) has become a re-emerging pathogen in cereal crops in the Czech Republic. WSMV was first reported in the former Czechoslovakia in the early 1980s, and then no record of the virus was documented until The incidence of the virus was recorded in recent years in several winter wheat field and many grass species. Here, we surveyed the incidence of WSMV in cereal crops. The results demonstrated the existence of the virus in winter wheat and volunteer wheat during each year of the monitoring period, which spanned from Although the range of infected samples was low (6.4% of the total tested samples), a high incidence of well-distributed virus was recorded. In at least six fields, the virus reached severe and potentially epidemic levels. In accordance with our previous report detailing WSMV infection of native grasses, we tested several grass species commonly grown in the Czech Republic. We found that some grass species acted as experimental hosts and possible reservoirs of the virus; these included Anthoxanthum odoratum (sweet vernal grass), Arrhenatherum elatius (false oat-grass), Lolium multiflorum (Italian rye-grass), Bromus japonicus (Japanese chess), Echinochloa crus-galli (barnyard grass), Holcus lanatus (meadow soft grass) and Holcus mollis (creeping soft grass). Some of these grass species are also important weeds of cereals, which may be the potential source of WSMV infection in cereal crops. Several widely used winter wheat cultivars were tested in the field after artificial inoculation with WSMV to evaluate virus titre by RT-qPCR. Overall, the tested cultivars had a low virus titre, which is associated with mild disease symptoms and may provide a good level of crop resistance to WSMV. Keywords: WSMV, survey, grass species, cereal crops 1. Introduction Received 14 July, 2016 Accepted 19 September, 2016 Correspondence Jiban Kumar Kundu, Tel: , Fax: , , CAAS. All rights reserved. Published by Elsevier Ltd. doi: /S (16) Wheat streak mosaic virus (WSMV) is the type member of the genus Tritimovirus of the family Potyviridae (Rabenstein et al. 2004). WSMV is widespread throughout the world and represents a severe threat in most wheat-growing regions (Rabenstein et al. 2002; Hadi et al. 2011; Coutts et al. 2014). WSMV infects many plant species of the family Poaceae (Brakke 1971; French and Stenger 2002), including wheat (Triticum aestivum L.), oat (Avena sativa L.), barley (Hor-

2 *** et al. Journal of Integrative Agriculture 2017, 16(0): deum vulgare L.), maize (Zea mays L.), millet (Panicum, Setaria, and Echinochloa spp.) and several grasses (see Table 1). Symptoms caused by WSMV include leaf mottling (mosaic pattern of green and chlorotic zones) and leaf streaking and are mainly observed in wheat (Vacke et al. 1986; Murray et al. 1998). The symptoms may progress to chlorosis and severe stunting of the plant, and in many cases, plants become sterile or produce shrivelled seeds (Ellis et al. 2004). WSMV is transmitted by wheat curl mites (WCMs, Aceria tosichella) (Slykhuis 1955; Orlob 1966), which are dispersed passively by air currents (Slykhuis 1955). The virus is also transmitted at low levels by seeds in wheat (Jones et al. 2005). Estimated levels of WSMV in seed lots originating from infected commercial wheat crops range from 0 to 0.22%, with an overall transmission rate of 0.06% (Lanoiselet et al. 2008). WSMV was first reported in the Great Plains Region (GPR) of North America in Since then, WSMV has been recorded in many wheat-growing regions of the world, including North and South America, Europe, the Middle East, Asia, Australia and New Zealand (Navia et al. 2013; Skoasracka et al. 2014). Crop losses caused by WSMV vary widely, ranging from 7 to 13% in Kansas (Atkinson and Grant 1967), reaching 18% in Canada (Christian and Willis 1993) and reaching up to 83% in wheat in Australia (Lanoiselet et al. 2008). WSMV spreads to cereal crops from virus reservoirs, which can include volunteer wheat plants and some grasses. At least 45 grass species are reported to be natural hosts for WSMV, and the majority of them are annuals (Sill and Agusiobo 1955; Christian and Willis 1993; French and Stenger 2002; Ito et al. 2012). Many of these WSMV grass hosts are also hosts for WCM (for review, see Navia et al. 2013). There is limited protection of cereal crops against WSMV and its WCM vector. Host resistance against the virus and/ or vector is the most effective way to reduce the yield loss caused by WSMV (Thomas et al. 2004; Richardson et al. 2014). Three resistance genes, known as Wsm1, Wsm2, and Wsm3, have been identified and introduced in wheat breeding lines. Wsm1 and Wsm2 resistance sources were found to be more temperature-sensitive (Seifers et al. 2013) than Wsm3 (Fahim et al. 2012); however, they have not yet been introduced into a commercial cultivar. The Wsm2 gene is the most widely used and has been successfully introduced into several wheat cultivars, including RonL (Seifers et al. 2006), Snowmass (Haley et al. 2011), Clara CL (Martin et al. 2014) and Oakley CL (Zhang et al. 2015a). However, with the deployment of WSMV-resistant cultivars, the limited resistance sources in these cultivars may be broken down by selective pressure on the virus (Zhang et al. 2015b). WSMV was first reported in the former Czechoslovakia in the early 1980s (Vacke et al. 1986). The incidence of the virus in recent years has become more frequent in winter wheat crops (Gadiou et al. 2009; Dráb et al. 2015). Vacke et al. (1986) reported the existence of WSMV in wheat, barley, oat and some cultivars of corn as well as in some grass species, including Avena fatua, Avena strigesa, Panicum miliaceum, Lolium multiflorum, Lagurus ovatus and Digitaria sanguinalis. In the present work, we report the results of a four-year survey of the incidence of WSMV in cereal crops, which was recorded for several winter wheat fields each year. We previously found that several grass species can become naturally infected with WSMV (Dráb et al. 2014), which may be a source of virus inoculum for cereal crops. We tested several grass species commonly grown in the Czech Republic and found that some of them were experimental hosts and possible reservoirs of the virus, such as Anthoxanthum odoratum (sweet vernal grass), Arrhenatherum elatius (false oat-grass), L. multiflorum (Italian rye-grass), Bromus japonicus (Japanese chess), Echinochloa crus-galli (barnyard grass), Holcus lanatus (meadow soft grass) and Holcus mollis (creeping soft grass). Several widely used winter wheat cultivars were tested in field experiments after artificial inoculation with WSMV to evaluate virus titre using real-time RT-qPCR. Several of the wheat cultivars, including Caldwell, Cubus, Florida, Matchball and Cim rana, had low levels of virus uptake. Furthermore, in one triticale cultivar, Koler, WSMV was undetected even when highly sensitive RT-qPCR was used. This low virus uptake in cultivars may be associated with a significant level of crop resistance to WSMV. 2. Materials and methods 2.1. Survey of WSMV in crop and grass hosts Samples from cereal fields, the fields margins and perennial meadows were collected from in different regions of the Czech Republic. The samples were collected during the early spring to autumn period randomly based on disease suspicion in the fields (Table 2) Experimental hosts within grass species Twenty-four species of grass (see Table 2) from Poaceae that naturally occur in the agro-ecosystem of the Czech Republic were tested. The plants were grown in greenhouses, and they were mechanically inoculated with a WSMV isolate (CZlab, accession number FJ216408) at the two-leaf stage. To accomplish this, individual plant leaves were smeared with sap containing the WSMV isolate with 0.1 mol L 1 phosphate buffer (ph 7.2) and carborundum powder. One month post-inoculation, the plants were tested by TAS-ELISA and then by RT-PCR as described in Gadiou et al. (2009).

3 4 *** et al. Journal of Integrative Agriculture 2017, 16(0): Table 1 Host species of Wheat streak mosaic virus (WSMV) Host Common name References Agropyron repens Couch grass Dráb et al Agrostis capillaris Common Bent In this study Alopecurus pratensis Meadow foxtail Dráb et al Anthoxanthum odoratum Sweet vernal-grass In this study Arrhenatherum elatius False oat-grass Dráb et al Austrostipa compressa Speargrass Vincent et al Aegilops cylindrica Jointed goatgrass Sill et al Avena barbata Bearded oat Coutts et al Avena fatua Wild oat Vacke et al Avena sativa Oat Brakke 1971 Avena strigesa Wild oats Vacke et al Avena sterilis Wild oats Murray et al Briza maxima Blowfly grass Coutts et al Bromus arvenis Field brone Sill et al Bromus diandrus Great brome Murray et al Bromus japonicus Japanese brome Wegulo et al Bromus rigidus Brome grass Coutts et al Bromus secalinus Cheat grass Sill et al Bromus tectorum Downy brome Sill et al Cynodon dactylon Couch grass Ellis et al Cenchrus longispinus Mat sandbur Connin 1956 Cenchrus pauciflours Sandbur Wegulo et al Digitaria sanguinalis Hairy crab grass Vacke et al. 1986; Somensen and Sill 1970 Echinochloa crus-galli Barnyardgrass Sill and Connin 1953 Echinochloa colonum Junglerice Khadivar and Nasrolahnejad 2009 Elymus repens Quackgrass Ito et al Eragrostis cilianensis Stink grass Connin 1956 Eragrostis curvula African lovegrass Ellis et al Eriochloa acuminata Tapertip cupgrass Seifers et al Eriochloa contracta Prairie cupgrass Christian and Willis 1993 Eleusine indica Crowsfoot Murray et al Eleusine tristachya Spike goosegrass Ellis et al Elymus canadensis Canada wild rye Ito et al Holcus lanatus Soft-grass In this study Holcus mollis Creeping soft grass In this study Hordeum leporinum Barley grass Coutts et al Hordeum vulgare Barley Brakke 1971 Lagurus ovatus Hare's-tail Vacke et al Lolium mitiflorum Annual ryegrass Vacke et al. 1986; Ellis et al Lolium rigidum Ryegrass Murray et al. 2005; Coutts et al Panicum dichotomiflorum Fall panicgrass Sill and Connin 1953 Panicum capillare Witch grass Coutts et al. 2008a, b Panicum millaceum Broomcorn millet Sill and Agusiobo 1955; Vacke et al. 1986; Ellis et al Pennisetum glaucum Pearl millet Seifers et al Phalaris aquatica phalaris Ellis et al Phleum pratense Timothy-grass Dráb et al Poa pratensis Bluegrass Ito et al. 2012; Dráb et al Secale cereale Cereal rye Vacke et al. 1986; Ito et al Setaria verticellata Whorled pigeon grass Murray et al Setaria viridis Green bristlegrass Sill and Connin 1953 Setaria italica Foxtail millet Truol et al Sorghum bicolor Sorgum Seifers et al Tragus australianus Small burr grass Coutts et al. 2008a, b Triticum aestivum Wheat Brakke 1971 Zea mays Maize Brakke 1971

4 *** et al. Journal of Integrative Agriculture 2017, 16(0): Table 2 Survey of the incidence of WSMV in wheat and barley crops and volunteer plants Sampling year Crop species Total tested samples Number of sampling fields Number of WSMV-positive samples Number of MSMV-infected field Percentage of WSMV-infected samples (%) Percentage of WSMV-infected fields (%) 2013 Winter wheat Winter barley Spring barley Winter wheat Winter barley Winter wheat Winter barley Spring barley Wheat volunteer Winter wheat Winter barley Spring barley Total Detection of WSMV TAS-ELISA WSMV was detected by triple anitbody sandwich (TAS)-ELISA using commercial antiserum (SEDIAG SAS, France) according to the manufacturer s instructions. RNA isolation WSMV-infected leaves were used for total RNA extraction using a Spectrum Plant Total RNA Kit (Sigma Aldrich, USA) following the manufacturer s instructions with minor modifications. A total of 100 mg of infected leaves was homogenized in liquid nitrogen to obtain a very fine powder. 500 µl of lysis buffer supplemented with 2-mercaptoethanol were added and mixed vigorously. The samples were incubated at 60 C for 5 min and centrifuged at r min 1 for 5 min. The supernatant was transferred to a filtration column and centrifuged again. 500 µl of binding solution were added to the filtrate, followed by centrifugation for 1 min at r min 1. The column was washed with 500 μl of wash buffer I and centrifuged. The column was washed three times with 500 μl of wash buffer II before adding 50 μl of elution buffer. The RNA was quantified using a Nanodrop 2000 (Thermo Scientific, USA) and stored at 80 C for further use. RNA was treated with DNase I using DNA-free (Ambion, USA) RT-PCR cdna synthesis First-strand cdna was synthesised using a Reverse Transcription System (Promega, USA). A reaction mixture composed of 1 µg RNA and 0.5 µg random primer was incubated at 70 C for 5 min and chilled on ice for 2 min. Then, the following components were added in the given order: 5 reaction buffer, 20 U µl 1 RNasin RNase inhibitor, 10 mmol L 1 dntp mix and 200 U µl 1 M-MLV reverse transcriptase enzyme. The mixture was incubated at 37 C for 1 h. The reaction was stopped by heating at 70 C for 10 min and subsequent chilling on ice. PCR cdna was amplified as described in Gadiou nt et al. (2009) using the primer pair WSMspeFw (5 - GCCTCGACACGGGAGCTA nt 3 ) and WSMspeRv ( nt ACCCATCCAGGAAGCAAGG nt -3 ). The reaction mixture contained cdna, 10 μmol L 1 each of forward and reverse primers, and 2 DreamTaq Green PCR Master Mix (Thermo Scientific, USA). The PCR conditions were as follows: initial denaturation at 94 C for 5 min, followed by 30 amplification cycles (94 C, 45 s; 66 C, 30 s; and 72 C, 1 min), and a final extension at 72 C for 10 min. The PCR assay yielded a 358-bp product. The PCR product was then separated on a 1% agarose gel and stained with SYBR Safe DNA Gel Stain (Invitrogen, USA) Quantitative analysis of WSMV in winter wheat cultivars Plant material and field experiment Quantitative analysis of WSMV was performed in wheat cultivars, including Florida, Vlada, Alcedo, Caldwell, Rocky, Ludwig, Cubus, Manager, Muszelka, Cim rana, Tobak, Matchball, Genius, Asano, and Bohemia as well as in the triticale cultivar Kolor. These cultivars are commonly grown in the Czech Republic. Each cultivar was grown in two rows (total ten plants per row) and in two replicates, one for virus inoculation and another for a non-inoculated control. The plants were mechanically inoculated with a WSMV isolate (CZlab, accession number FJ216408) at the three-leaf stage during autumn as described above. The leaf samples of the tested cultivars were collected in the spring at Feekes growth stage 9 for WSMV titre analysis by RT-qPCR. RT-qPCR RNA was isolated as described above. qpcr was performed as described in Dráb et al. (2014) using primer pair WSMV-F ( nt AAGTGCAGAACAGCGTTG 9

5 6 *** et al. Journal of Integrative Agriculture 2017, 16(0): nt -3 )/WSMV-R ( nt AAACTGTGCGTGTTCTCC nt - 3 ) (Tatineni et al. 2010). A primer pair for wheat elongation factor 1α (eef1α) served as a positive control, enabling the assessment of RNA quality (Jarošová and Kundu 2010). The qpcr reaction was performed on a LightCycler 480 Real-Time PCR System (Switzerland). A 250 nmol L 1 primer concentration was used in the qpcr reaction mixture, and the primer annealing temperature was 60 C. The total cdna concentration in the reaction mixture was 3 ng ml 1. The amplification protocol was as follows: denaturation at 95 C for 10 min, followed by 40 cycles of 95 C for 5 s, 60 C for 30 s and 72 C for 10 s. Finally, melting analysis of the PCR products was performed. qpcr data analysis The titre of WSMV in the tested samples was calculated according to a ten-fold standard dilution curve constructed using the PCR product of a positive WSMV sample. Calculations were made using LightCycler 480 Real-Time PCR System software. The PCR product quantity in µg was converted to pmol using the average molecular weight of a ribonucleotide (340 Da) and the number of bases of the transcript (Nb). The following mathematical formula was applied: pmol of dsdna=μg of dsdna) 10 6 pg μg 1 /660 pmol pg -1 /Nb. Avogadro s constant ( molecules mol 1 ) was used to estimate the number of transcripts. A graphical representation of the data was constructed using the programme Microsoft Excel. 3. Results 3.1. Incidence of WSMV in crops were tested after mechanical inoculation with WSMV. The virus was detected in eight species, including Agrostis capillaris, A. odoratum, L. multiflorum, B. japonicas, A. elatius, E. crus-galli, H. lanatus and H. mollis (Table 3 and Fig. 2). WSMV was generally detectable by both TAS-ELISA and RT-PCR, except for in A. capillaris and H. lanatus, in which WSMV was detected solely by RT-PCR Differences in virus uptake among wheat cultivars WSMV uptake in commonly grown winter wheat cultivars was tested by analysis of virus titre by RT-qPCR. Ten cul- Fig. 1 Map of Wheat streak mosaic virus (WSMV) occurrence in the fields of the Czech Republic. WSMV infections are indicated as stars: black stars, fields with individual infected plants; red stars, fields with severe infections. WSMV incidence was surveyed from the years 2013 to Sampling was performed in a random manner in a total of 134 fields; the fields included winter wheat and barley, spring barley and volunteer wheat plants. In total, 876 samples were tested, and the infection rate varied among years and crops. WSMV was detected solely in winter wheat, in both crop and voluntary plants. The average rate of WS- MV-infected samples was 6.4% (56 samples) in total, and among the 94 wheat fields and four wheat volunteer fields tested, the virus was detected in 19 wheat fields and one wheat volunteer field (Table 2). These results indicate that only a few individual infected plants were detected in most of the fields. However, in six fields, the virus was found at a high incidence and was widely distributed; these included two fields sampled in 2013, one field sampled in 2015 and three fields sampled in 2016 (the locations of these fields are indicated in red on the map in Fig. 1) Experimental hosts within the grass species Nineteen commonly grown grass species, including weeds, Table 3 Detection of WSMV in grass species after artificial inoculation with WSMV Number of WSMV-positive plants/ Grass species Number of tested plants TAS-ELISA RT-PCR Aera spica-venti 0/2 0/2 A. capillaris 0/7 2/7 Agrostis stolonifera 0/7 0/7 A. odoratum 1/7 1/7 A. elatius 2/7 2/7 Bromus erectus 0/4 0/4 B. japonicus 4/7 4/7 Bromus sterilis 0/4 0/4 Cynosurus cristatus 0/7 0/7 Dactylis glomerata 0/5 0/5 E. crus-galli 5/7 5/7 Holcus lanatus 0/5 1/5 H. mollis 1/5 1/5 Hordeum murinum 0/5 0/5 L. multiflorum 2/6 2/6 P. pratense 0/7 0/7 Poa annua 0/5 0/5 Poa trivialis 0/6 0/6 Trisetum flavescens 0/7 0/7

6 *** et al. Journal of Integrative Agriculture 2017, 16(0): tivars were tested in field experiments, including a triticale cultivar. Significant differences in virus titre were detected in the tested cultivars (Fig. 3). The WSMV uptake levels that were recorded in the tested cultivars were grouped as follows: I) low: Caldwell, Cubus, Florida, Matchball and Cim rana; II) medium: Rocky, Vláda, Alcedo, Muszelka and Manager; and III) high: Ludwig, Bohemia, Asano, Tobak and Genius. Notably, WSMV was undetectable in the triticale cultivar Koler (Fig. 3). 4. Discussion The results of the current survey of the incidence of Wheat streak mosaic virus (WSMV) in the Czech Republic showed a low occurrence of the virus and demonstrated that the virus was present only in wheat. Winter wheat is the main host of WSMV and the most commonly cultivated cereal crop in the Czech Republic. In our study, the majority of tested samples came from wheat crops rather than from minor barley, in which WSMV was not detected (see Table 2). 358 bp M Fig. 2 Electrophoresis of RT-PCR products for the detection of WSMV in grass species after artificial inoculation with WSMV. SYBR Green-stained 1% agarose gel showing a PCR product of 358 bp. Lanes 1 19, A. spica-venti, A. odoratum, T. flavescens, L. multiflorum, D. glomerata, C. cristatus, B. japonicus, B. sterilis, B. erectus, A. elatius, A. pratensis, A. capillaris, A. stolonifera, E. crus-galli, P. annua, P. trivialis, H. lanatus, H. mollis, and H. murinum; lane M, GeneRuler 100 bp Plus DNA Ladder (Thermo Scientific, USA). We previously found that WSMV occurs very occasionally and that only individual plants within a field are affected in general. In at least six winter wheat fields (early sowing), we found a high incidence of WSMV, and the virus was spread throughout the fields (see the WSMV symptoms in wheat cultivars in Fig. 4-A nad B). These results suggest that there is significant potential for WSMV infection to increase in crops. The warmer autumns and winters experienced by Central Europe in recent years have created favourable preconditions for the life cycle of the virus vector, namely, wheat curl mites (WCMs). WCMs have been reported to survive for months at near-freezing temperatures, including for several days at approximately 18 C (Townsend and Johnson 1996). No report on the occurrence of WCMs in the Czech Republic is currently available, and several attempts to identify WCMs in infected wheat crops were unsuccessful in our hands. However, WCMs have been reported to be abundant in neighbouring countries (Skoracka et al. 2014). The incidence of WSMV in natural grass species (Dráb et al. 2014) indicates the presence of the vector in the agro-ecosystem and thereby the introduction of the virus into crops. Another possibility for WSMV introduction is through infected seed, which may have occurred at a low level; this type of dissemination of the virus was reported in New Zealand (Lebas et al. 2009). The transmission rate of WSMV via seed ranges up to 1.5% (Jones et al. 2005), and seeds are likely an important source of virus inoculum when WCMs are also present (Lanoiselet et al. 2008). WSMV may cause significant yield losses in wheat, which may reach over 90% (Hadi et al. 2011) depending on weather conditions, cultivars and growth stage affected by virus infection. Generally, infections during the early stages of plant growth result in higher yield losses (Hunger et al. 1992; Baley et al. 2001). WSMV infections are also associated with reductions in root biomass and the efficiency of water uptake (Price WSMV copy number 1.40E E E E E E E E+00 WSMV titre analysis by RT-qPCR Kolor Caldwell Cubus Florida Matchball Cim rana Rocky Vláda Alcedo Muszelka Manager Ludwig Bohemia Asano Tobak Genius Cultivar Fig. 3 WSMV titre (copy number) in wheat cultivars and triticale cultivar Koler. One sample from each cultivar was tested.

7 8 *** et al. Journal of Integrative Agriculture 2017, 16(0): A B Fig. 4 WSMV symptoms in a naturally infected field growing wheat cultivars Hymack (A) Bodyček (B). Symptoms appear as small chlorotic lines, yellow and green streaks and a mosaic pattern in the leaves. The symptomatic leaves later become necrotic and dried out. et al. 2010), creating serious concerns with regard to climate change. In addition to crops, grass species are major hosts for both WSMV and WCMs, and a good number of them are common hosts (reviewed in Navia et al. 2013). In the Czech Republic, we observed the natural occurrence of WSMV in several grass species, including E. repens, A. pratensis, A. elatius, P. pratense and P. pratensis (Dráb et al. 2014). Several other species, including A. fatua, D. sanguinalis, L. multiflorum and P. miliaceum, were also reported by Vacke et al. (1986). Hence, grass species can serve as reservoirs of the virus and a source of virus introduction into field crops. The grass host species of WSMV are shown in Table 1. In the present report, we describe five species, namely, A. capillaris, A. odoratum, B. sterilis, H. lanatus and H. mollis, as new hosts of WSMV, and several of the other species identified (L. multiflorum, B. japonicus, A. elatius and E. crus-galli) were confirmed to be hosts in earlier reports (Vacke et al. 1986; Coutts et al. 2008b). The broad host range of WSMV and WCMs among Poaceae species creates the potential threat of virus invasion in cereal crops, in particular winter wheat. Some of these grass species are important weeds of cereal crops, including E. crus-galli, E. repens, L. multiflorum, B. japonicas, and P. pratensis. These weeds may act as reservoirs for WSMV, facilitating its introduction into wheat crops. In our survey, WSMV was detected solely in winter wheat, and its incidence was recorded as approximately 16% in the tested field. However, no evidence exists for the Czech Republic regarding whether such restriction of the virus in crops occurs through seed transmission, as was reported in New Zealand (Lebas et al. 2009). Host resistance to WSMV infection might be an effective method of disease control. The sources of resistance to WSMV are temperature-sensitive (Fahim et al. 2012). Cultivars derived from Wsm2, including RonL (Seifers et al. 2007), Snowmass (Haley et al. 2011), Clara CL (Martin et al. 2014) and Oakley CL (Zhang et al. 2015a), were reported to be resistant to WSMV isolates from the USA. The cultivars Mace, Millennium and Pronghorn were shown to be cultivated to a certain level of tolerance to WSMV in Nebraska, USA (Wegulo et al. 2008). Thus far, no evidence of resistance in commonly grown wheat cultivars in Central Europe is available. Quantitative analysis of virus titre using real-time qpcr (Balaji et al. 2003) seems to be very effective for characterising the level of resistance and provides significant enhancement in addition to biological assays (Jarošová et al. 2013). Field experiments involving mechanical inoculation of WSMV showed a low level of WSMV uptake in some winter cultivars, such as Caldwell, Cubus, Florida, Matchball and Cim rana. We previously showed that virus titre (copy number) is closely correlated with a high level of cereal (wheat and barley) resistance to Barley yellow dwarf virus (Jarošová et al. 2013). The low WSMV uptake in the above-mentioned cultivars may also be associated with potentially significant resistance to the virus under field conditions (Fig. 4). However, for more thorough evaluation of the resistance of the tested cultivars, multiple traits and characteristics need to be considered. Furthermore, host resistance to WSMV in wheat has been reported to be temperature-sensitive (Seifers et al. 2013), which is a major concern for the development of new cultivars with stable resistance against virus-host interactions under field conditions. Hence, environmental conditions (i.e., temperature) during the growing session play a significant role in the stability of wheat resistance to WSMV in the field. 5. Conclusion Our results provide significant evidence that the presence of WSMV in winter wheat crops, together with Wheat dwarf virus and Barley yellow dwarf virus, may cause serious problems with respect to crop protection. Grass species, in particular the weed hosts described here, act as reservoirs of WSMV and are possibly the key source of the introduction of WSMV into winter wheat (cereal crops). Quantitative analysis of virus uptake in wheat cultivars and one cultivar of triticale showed that some cultivars, including Caldwell, Cubus, Florida, Matchball and Cim rana, had a low virus titre. Our results suggest that the low virus titre detected in these cultivars may be associated with a significant level of resistance to WSMV under field conditions. However, further analysis based on multiple resistance traits associated with yield loss should be performed to more thoroughly evaluate the resistance status of the cultivars. Acknowledgements We thank Mrs. Zuzana Červená (Division of Crop Protection and Plant Health, Crop Research Institute, Czech Republic) for her technical assistance and Dr. Fousek and Dr. Neubau-

8 *** et al. Journal of Integrative Agriculture 2017, 16(0): erová (Division of Crop Protection and Plant Health, Crop Research Institute, Czech Republic) for their help with some experiments. This work was supported by the grants from the Ministry of Agriculture, Czech Republic from projects QJ (50%) and RO0415 (50%). References Atkinson T G, Grant M N An evaluation of streak mosaic losses in winter wheat. Phytopathology, 57, Balaji B, Bucholtz D B, Anderson J M Barley yellow dwarf virus and Cereal yellow dwarf virus quantification by realtime polymerase chain reaction in resistant and susceptible plants. Phytopathology, 93, Baley G J, Talbert L E, Martin J M, Young M J, Habernicht D K, Kushnak G D, Berg J E, Lanning S P, Bruckner P L Agronomic and end-use qualities of Wheat streak mosaic virus resistant spring wheat. Crop Science, 41, Brakke M K Wheat streak mosaic virus. CMI/AAB Descriptions of Plant Viruses. No. 48. Association of Applied Biologists, Wellesbourne, UK. showdpv.php?dpv no=48 Christian M L, Willis W G Survival of Wheat streak mosaic virus in grass hosts in Kansas from what harvest to fall wheat emergence. Plant Disease, 77, Connin R V The host range of the wheat curl mite, vector of wheat streak mosaic. Journal of Economic Entomology, 48, 1 4. Coutts B A, Banovic M, Kehoe M A, Severtson D L, Jones R A C Epidemiology of Wheat streak mosaic virus in wheat in a Mediterranean-type environment. European Journal of Plant Pathology, 140, Coutts B A, Hammond N E B, Kehoe M A, Jones R A C. 2008a. Finding Wheat streak mosaic virus in southwest Australia. Australian Journal of Agricultural Research, 59, Coutts B A, Strickland G R, Kehoe M A, Severtson D L, Jones R A C. 2008b. The epidemiology of Wheat streak mosaic virus in Australia: Case histories, gradients, mite vectors, and alternative hosts. Australian Journal of Agricultural Research, 59, Dráb T, Svobodová E, Ripl J, Jarošová J, Rabenstein F, Melcher U, Kundu J K SYBR Green I based RT-qPCR assays for the detection of RNA viruses of cereals and grasses. Crop and Pasture Science, 65, Dráb T, Svobodová E, Ripl J, Slavíková L, Kumar J Reservoirs of viral pathogens in wild grasses. Úroda, 63, (in Czech) Ellis M H, Rebetzke G J, Kelman W M, Moore C S, Hyles J E Detection of Wheat streak mosaic virus in four pasture grass species in Australia. Plant Pathology, 53, 239. Fahim M, Larkin P J, Haber S, Shorter S, Lonergan P F, Rosewarne G M Effectiveness of three potential sources of resistance in wheat against Wheat streak mosaic virus under field conditions. Australas Plant Pathology, 41, French R, Stenger D C Wheat streak mosaic virus. CMI/ AAB Descriptions of Plant Viruses. No Association of Applied Biologists, Wellesbourne, UK. net/dpv/showdpv.php?dpv no=393 Gadiou S, Kúdela O, Ripl J, Rabenstein F, Kundu J K, Glasa M An amino acid deletion in Wheat streak mosaic virus capsid protein distinguishes a homogeneous group of European isolates and facilitates their specific detection. Plant Disease, 93, Hadi B A R, Langham M A C, Osborne L, Tilmon K J Wheat streak mosaic virus on wheat: Biology and management. Journal of Integrated Pest Management, 2, doi: /IPM10017 Haley S D, Johnson J J, Peairs F B, Stromberger J A, Heaton E E, Seifert S A, Kottke R A, Rudolph J B, Martin T J, Bai G, Chen X, Bowden R, Jin Y, Kolmer J A, Seifers D L, Chen M, Seabourn B W Registration of Snowmass wheat. Journal of Plant Registrations, 5, Hunger R M, Sherwood J L, Evans C K, Montana J R Effects of planting date and inoculation date on severity of wheat streak mosaic in hard red winter wheat cultivars. Plant Disease, 76, Ito D, Miller Z, Menalled F, Moffet M, Burrows M Relative susceptibility among alternative host species prevalent in the Great Plains to Wheat streak mosaic virus. Plant Disease, 96, Jarošová J, Chrpová J, Šíp V, Kundu J K A comparative study of the Barley yellow dwarf virus species PAV and PAS: Distribution, accumulation and host resistance. Plant Pathology, 62, Jarošová J, Kundu J K Validation of reference genes as internal control for studying viral infections in cereals by quantitative real-time PCR. BMC Plant Biology, 10, 146. Jones R A C, Coutts B A, Mackie A E, Dwyer G I Seed transmission of Wheat streak mosaic virus shown unequivocally in wheat. Plant Disease, 89, Khadivar R S, Nasrolahnejad S Serological and molecular detection of Wheat streak mosaic virus (WSMV) in cereal fields of Golestan province, Northern Iran. J Plant Prod, 16, 4 Lebas B S M, Ochoa-Corona F M, Alexander B J R, Lister R A, Fletcher J D F, Bithel S L l, Burnip G M First report of Wheat streak mosaic virus on wheat in New Zealand. Plant Disease, 93, 430. Lanoiselet V M, Hind-Lanoiselet T L, Murray G M Studies on the seed transmission of Wheat streak mosaic virus. Australas Plant Pathology, 37, Martin T J, Zhang G, Fritz A K, Miller R, Chen M Registration of Clara CL Wheat. Journal of Plant Registrations, 8, Murray T D, Parry D W, Cattlin N D A Colour Handbook of Diseases of Small Grain Cereal Crops. Manson Publishing, London. Murray G M, Knihinicki D K, Wratten K, Edwards J Wheat Streak Mosaic and the Wheat Curl Mite. NSW Department of Primary Industries, Orange NSW Australia. Primefact 99.

9 10 *** et al. Journal of Integrative Agriculture 2017, 16(0): Navia D, de Mendonca R S, Skoracka A, Szydlo W, Knihinicki D, Hein G L, da Silva Pereira P R V, Truol G, Lau D Wheat curl mite, Aceria tosichella, and transmitted viruses: An expanding pest complex affecting cereal crops. Experimental and Applied Acarology, 59, Orlob G Feeding and transmission characteristics of Aceria tulipae Keifer as a vector of Wheat streak mosaic virus. Phytopathol Z, 55, Price J A, Workneh F, Evett S R, Jones D C, Arthur J, Rush C M Effects of Wheat streak mosaic virus on root development and water-use efficiency of hard red winter wheat. Plant Disease, 94, Rabenstein F, Stenger D C, French R Genus tritimovirus. In: Lapierre H, Signoret P A, eds., Viruses and Virus Diseases of Poaceae (Gramineae). INRA, France. pp Rabenstein F, Seifers D L, Schubert J, French R, Stenger D C Phylogenetic relationships, strain diversity and biogeography of tritimoviruses. Journal of General Virology, 83, Richardson K, Miller A D, Hoffmann A A, Larkin P Potential new sources of wheat curl mite resistence in wheat to prevent the spread of yield-reducing pathogens. Experimental and Applied Acarology, 64, Seifers D L, Harvey T L, Kofoid K D, Stegmeier W D Natural infection of pearl millet and sorghum by Wheat streak mosaic virus in Kansas. Plant Disease, 80, Seifers D L, Martin T J, Harvey T L, Haber S Temperature sensitive Wheat streak mosaic virus resistance identified in KS03HW12 wheat. Plant Disease, 91, Seifers D L, Martin T J, Harvey T L, Haber S, Haley S D Temperature sensitive and efficacy of Wheat streak mosaic virus resistance derived from CO wheat. Plant Disease, 90, Seifers D L, Haber S, Martin T J, Zhang G New sources of temperature sensitive resistance to Wheat streak mosaic virus in wheat. Plant Disease, 97, Sill Jr W H, Connin R V Summary of the known host range of Wheat streak mosaic virus. Transactions of the Kansas Academy of Science, 56, Somensen H W, Sill Jr W H The wheat curl mite, Aceria tulipae Keifer, in relation to epidemiology and control of wheat streak mosaic. Kansas State University, Agricultural Experiment Station, Research Publication, 162, Sill Jr W H, Agusiobo P C Host range studies of the Wheat streak mosaic virus. Transactions of the Kansas Academy of Science, 56, Skoracka A, Rector B, Kuczyński L, Szydło W, Hein G, French R Global spread of wheat curl mite by its most polyphagous and pestiferous lineages. Annals of Applied Biology, 165, Slykhuis J T Aceria tulipae keifer (Acarina: Eriophyidae) in relation to the spread of wheat streak mosaic. Phytopathology, 45, Tatineni S, Graybosch R A, Hein G L, Wegulo S N, French R Wheat cultivar-specific disease synergism and alteration of virus accumulation during co-infection with Wheat streak mosaic virus and Triticum mosaic virus. Phytopathology, 100, Thomas J B, Conner R L, Graf R J Comparison of different sources of vector resistance for controlling wheat streak mosaic in winter wheat. Crop Science, 44, Townsend L, Johnson D Wheat streak mosaic virus and the wheat curl mite. University of Kentucky, ENTFACT-117. [ ]. ef117.asp Truol G, Sagadin M, Rodriguez M Fox tail millet (Setaria italica L.): a new reservoir species of the Wheat streak mosaic virus (WSMV) in the province of Buenos Aires. Biocell, 34, A135. Vacke J, Zacha V, Jokeš M Identification of virus in wheat new to Czechoslovakia. In: Proceeding X Czechoslovak Plant Protection. Conference, Brno. pp Vincent S J, Coutts B A, Jones R A C Effects of introduced and indigenous viruses on native plants: Exploring their disease causing potential at the agro-ecological interface. PLOS ONE, 9, e Wegulo S N, Hein G L, Klein R N, French R C Managing Wheat Streak Mosaic. University of Nebraska, Lincoln (Extension EC1871). Zhang G, Martin T J, Fritz A K, Miller R, Chen M, Bowden R L, Johnson J. 2015a. Registration of Oakley CL wheat. Journal of Plant Registrations, 9, Zhang G, Seifers D L, Martin T J. 2015b. Inheritance of Wheat streak mosaic virus resistance in KS03HW12. Austin Journal of Plant Biology, 1, (Managing editor ZHANG Juan)

A Partial Host Range of the High Plains Virus of Corn and Wheat

A Partial Host Range of the High Plains Virus of Corn and Wheat A Partial Host Range of the High Plains Virus of Corn and Wheat Dallas L. Seifers, Associate Professor, Kansas State University, Agricultural Research Center-Hays, Hays 67601-9228; Tom L. Harvey, Professor,

More information

Diagnosis and Quantification of Strawberry Vein Banding Virus Using Molecular Approaches

Diagnosis and Quantification of Strawberry Vein Banding Virus Using Molecular Approaches Diagnosis and Quantification of Strawberry Vein Banding Virus Using Molecular Approaches Ali Mahmoudpour Department of Plant Pathology, University of California, Davis, CA, 95616, USA Current Address:

More information

Quant One Step RT-PCR Kit

Quant One Step RT-PCR Kit 1. Quant One Step RT-PCR Kit For fast and sensitive one-step RT-PCR www.tiangen.com/en RT121221 Quant One Step RT-PCR Kit Kit Contents Cat. no. KR113 Contents Hotmaster Taq Polymerase (2.5 U/μl) Quant

More information

Sensitivity vs Specificity

Sensitivity vs Specificity Viral Detection Animal Inoculation Culturing the Virus Definitive Length of time Serology Detecting antibodies to the infectious agent Detecting Viral Proteins Western Blot ELISA Detecting the Viral Genome

More information

TECHNICAL SHEET No. 21. Virus Detection: Potato virus Y (PVY)

TECHNICAL SHEET No. 21. Virus Detection: Potato virus Y (PVY) TECHNICAL SHEET No. 21 Virus Detection: Potato virus Y (PVY) Method: Immunocapture RT-PCR, RFLP Immunocapture RT-PCR General Virus detected: PVY from potato General method: IC-RT-PCR, RFLP-IC-RT-PCR Developed

More information

SuperScript IV Reverse Transcriptase as a better alternative to AMV-based enzymes

SuperScript IV Reverse Transcriptase as a better alternative to AMV-based enzymes WHITE PAPER SuperScript IV Reverse Transcriptase SuperScript IV Reverse Transcriptase as a better alternative to AMV-based enzymes Abstract Reverse transcriptases (RTs) from avian myeloblastosis virus

More information

HiPer RT-PCR Teaching Kit

HiPer RT-PCR Teaching Kit HiPer RT-PCR Teaching Kit Product Code: HTBM024 Number of experiments that can be performed: 5 Duration of Experiment: Protocol: 4 hours Agarose Gel Electrophoresis: 45 minutes Storage Instructions: The

More information

P HENIX. PHENIX PCR Enzyme Guide Tools For Life Science Discovery RESEARCH PRODUCTS

P HENIX. PHENIX PCR Enzyme Guide Tools For Life Science Discovery RESEARCH PRODUCTS PHENIX PCR Enzyme Guide PHENIX offers a broad line of premium quality PCR Enzymes. This PCR Enzyme Guide will help simplify your polymerase selection process. Each DNA Polymerase has different characteristics

More information

Identification of a Cucumber mosaic virus Subgroup II Strain Associated with Virus-like Symptoms on Hosta in Ohio

Identification of a Cucumber mosaic virus Subgroup II Strain Associated with Virus-like Symptoms on Hosta in Ohio 2013 Plant Management Network. Accepted for publication 18 December 2012. Published. Identification of a Cucumber mosaic virus Subgroup II Strain Associated with Virus-like Symptoms on Hosta in Ohio John

More information

Reverse transcription-pcr (rt-pcr) Dr. Hani Alhadrami

Reverse transcription-pcr (rt-pcr) Dr. Hani Alhadrami Reverse transcription-pcr (rt-pcr) Dr. Hani Alhadrami hanialhadrami@kau.edu.sa www.hanialhadrami.kau.edu.sa Overview Several techniques are available to detect and analyse RNA. Examples of these techniques

More information

INTRODUCTION TO REVERSE TRANSCRIPTION PCR (RT-PCR) ABCF 2016 BecA-ILRI Hub, Nairobi 21 st September 2016 Roger Pelle Principal Scientist

INTRODUCTION TO REVERSE TRANSCRIPTION PCR (RT-PCR) ABCF 2016 BecA-ILRI Hub, Nairobi 21 st September 2016 Roger Pelle Principal Scientist INTRODUCTION TO REVERSE TRANSCRIPTION PCR (RT-PCR) ABCF 2016 BecA-ILRI Hub, Nairobi 21 st September 2016 Roger Pelle Principal Scientist Objective of PCR To provide a solution to one of the most pressing

More information

AMV First Strand cdna Synthesis Kit

AMV First Strand cdna Synthesis Kit DNA AMPLIFICATION & PCR AMV First Strand cdna Synthesis Kit Instruction Manual NEB #E6550S Store at 20 C ISO 9001 Registered Quality Management ISO 14001 Registered Environmental Management ISO 13485 Registered

More information

ReverTra Ace qpcr RT Master Mix

ReverTra Ace qpcr RT Master Mix Instruction manual ReverTra Ace qpcr RT Master Mix 1203 F1173K ReverTra Ace qpcr RT Master Mix FSQ-201 200 reactions Store at -20 C Contents [1] Introduction [2] Components [3] Protocol 1. RNA template

More information

Technical Review. Real time PCR

Technical Review. Real time PCR Technical Review Real time PCR Normal PCR: Analyze with agarose gel Normal PCR vs Real time PCR Real-time PCR, also known as quantitative PCR (qpcr) or kinetic PCR Key feature: Used to amplify and simultaneously

More information

One Step SYBR PrimeScript RT-PCR Kit II (Perfect Real Time)

One Step SYBR PrimeScript RT-PCR Kit II (Perfect Real Time) Cat. # RR086A For Research Use One Step SYBR PrimeScript RT-PCR Kit II Product Manual Table of Contents I. Description...3 II. III. IV. Principle...3 Components...5 Storage...6 V. Features...6 VI. VII.

More information

SOP: SYBR Green-based real-time RT-PCR

SOP: SYBR Green-based real-time RT-PCR SOP: SYBR Green-based real-time RT-PCR By Richard Yu Research fellow Centre for Marine Environmental Research and Innovative Technology (MERIT) Department of Biology and Chemistry City University of Hong

More information

Human TNF qpcr primer pair

Human TNF qpcr primer pair Shipping and Storage information Catalog No. Amount Reaction number of 25-μl volume Shipping and Storage Package tube 2 nmol 200 lyophilized powder 1 Information of the target gene and primers Species

More information

Quantitation of mrna Using Real-Time Reverse Transcription PCR (RT-PCR)

Quantitation of mrna Using Real-Time Reverse Transcription PCR (RT-PCR) Quantitation of mrna Using Real-Time Reverse Transcription PCR (RT-PCR) Quantitative Real-Time RT-PCR Versus RT-PCR In Real-Time RT- PCR, DNA amplification monitored at each cycle but RT-PCR measures the

More information

Phosphate buffered saline (PBS) for washing the cells TE buffer (nuclease-free) ph 7.5 for resuspending the SingleShot RNA control template

Phosphate buffered saline (PBS) for washing the cells TE buffer (nuclease-free) ph 7.5 for resuspending the SingleShot RNA control template Catalog # Description 172-5085 SingleShot SYBR Green Kit, 100 x 50 µl reactions For research purposes only. Introduction The SingleShot SYBR Green Kit prepares genomic DNA (gdna) free RNA directly from

More information

ProtoScript. First Strand cdna Synthesis Kit DNA AMPLIFICATION & PCR. Instruction Manual. NEB #E6300S/L 30/150 reactions Version 2.

ProtoScript. First Strand cdna Synthesis Kit DNA AMPLIFICATION & PCR. Instruction Manual. NEB #E6300S/L 30/150 reactions Version 2. DNA AMPLIFICATION & PCR ProtoScript First Strand cdna Synthesis Kit Instruction Manual NEB #E6300S/L 30/150 reactions Version 2.2 11/16 be INSPIRED drive DISCOVERY stay GENUINE This product is intended

More information

mmu-mir-200a-3p Real-time RT-PCR Detection and U6 Calibration Kit User Manual MyBioSource.com Catalog # MBS826230

mmu-mir-200a-3p Real-time RT-PCR Detection and U6 Calibration Kit User Manual MyBioSource.com Catalog # MBS826230 mmu-mir-200a-3p Real-time RT-PCR Detection and U6 Calibration Kit User Manual Catalog # MBS826230 For the detection and quantification of mirnas mmu-mir-200a-3p normalized by U6 snrna using Real-time RT-PCR

More information

FOR RESEARCH USE ONLY. NOT FOR HUMAN OR DIAGNOSTIC USE.

FOR RESEARCH USE ONLY. NOT FOR HUMAN OR DIAGNOSTIC USE. Instruction manual RNA-direct SYBR Green Realtime PCR Master Mix 0810 F0930K RNA-direct SYBR Green Realtime PCR Master Mix Contents QRT-201T QRT-201 0.5mLx2 0.5mLx5 Store at -20 C, protected from light

More information

SYBR Green Realtime PCR Master Mix

SYBR Green Realtime PCR Master Mix Instruction manual SYBR Green Realtime PCR Master Mix 0810 F0924K SYBR Green Realtime PCR Master Mix QPK-201T 1 ml x 1 QPK-201 1 ml x 5 Contents [1] Introduction [2] Components [3] Primer design [4] Detection

More information

SideStep Lysis and Stabilization Buffer

SideStep Lysis and Stabilization Buffer SideStep Lysis and Stabilization Buffer INSTRUCTION MANUAL Catalog #400900 Revision B.0 For Research Use Only. Not for use in diagnostic procedures. 400900-12 LIMITED PRODUCT WARRANTY This warranty limits

More information

GeneCopoeia TM. All-in-One qpcr Mix For universal quantitative real-time PCR. User Manual

GeneCopoeia TM. All-in-One qpcr Mix For universal quantitative real-time PCR. User Manual GeneCopoeia TM Expressway to Discovery All-in-One qpcr Mix For universal quantitative real-time PCR Cat. No. AOPR-0200 (200 qpcr reactions) Cat. No. AOPR-0600 (600 qpcr reactions) Cat. No. AOPR-1000 (1000

More information

II First Strand cdna Synthesis Kit

II First Strand cdna Synthesis Kit DNA AMPLIFICATION & PCR ProtoScript II First Strand cdna Synthesis Kit Instruction Manual NEB #E6560S/L 30/150 reactions Version 1.5 12/17 be INSPIRED drive DISCOVERY stay GENUINE This product is intended

More information

mmu-mir-34a Real-time RT-PCR Detection Kit User Manual

mmu-mir-34a Real-time RT-PCR Detection Kit User Manual mmu-mir-34a Real-time RT-PCR Detection Kit User Manual Catalog # CPK1272 For the detection and quantification of mirna mmu-mir-34a using Real-Time RT-PCR detection instruments. For research use only. Not

More information

mirna Purification from Plant Tissues: Corn, Soybean and Arabidopsis

mirna Purification from Plant Tissues: Corn, Soybean and Arabidopsis mirna Purification from Plant Tissues: Corn, Soybean and Arabidopsis A Maxwell RSC mirna Tissue Kit Application Note Materials Required: Maxwell RSC mirna Tissue Kit (Cat.# AS1480) QuantiFluor RNA System

More information

MMLV Reverse Transcriptase 1st-Strand cdna Synthesis Kit

MMLV Reverse Transcriptase 1st-Strand cdna Synthesis Kit MMLV Reverse Transcriptase 1st-Strand cdna Synthesis Kit Cat. No. MM070150 Available exclusively thru Lucigen. lucigen.com/epibio www.lucigen.com MA265E MMLV Reverse Transcriptase 1st-Strand cdna Synthesis

More information

Molekulargenetische Reagenzien. Raumtemperaturstabile PCR und qpcr Reagenzien

Molekulargenetische Reagenzien. Raumtemperaturstabile PCR und qpcr Reagenzien Molekulargenetische Reagenzien Raumtemperaturstabile PCR und qpcr Reagenzien Polypeptide Stabilization Technology: Stability TAG Ice-free reaction set-up Our temperature stable enzymes allow you to change

More information

FMF NIRCA PROTOCOL STEP 1.

FMF NIRCA PROTOCOL STEP 1. FMF NIRCA PROTOCOL STEP 1. After you have isolated patient s DNA and DNA from a healthy donor (wild type), you perform a nested PCR. The primers used to amplify exon 2 and exon 10 of the mefv gene are

More information

RP RXN RTase/RI Enzyme Mix 5X RT Buffer (DTT/dNTPs) Oligo (dt)/random Primer Mix DEPC-Treated H2O

RP RXN RTase/RI Enzyme Mix 5X RT Buffer (DTT/dNTPs) Oligo (dt)/random Primer Mix DEPC-Treated H2O www.smobio.com Product Information Reverse Transcription Kit II RP1400 100 RXN RTase/RI Enzyme Mix 5X RT Buffer (DTT/dNTPs) Oligo (dt)/random Primer Mix DEPC-Treated H2O ExcelRT series 100 μl 500 μl 100

More information

T7-Based RNA Amplification Protocol (in progress)

T7-Based RNA Amplification Protocol (in progress) T7-Based RNA Amplification Protocol (in progress) Jacqueline Ann Lopez (modifications) Amy Cash & Justen Andrews INTRODUCTION T7 RNA Amplification, a technique originally developed in the laboratory of

More information

Percent survival. Supplementary fig. S3 A.

Percent survival. Supplementary fig. S3 A. Supplementary fig. S3 A. B. 100 Percent survival 80 60 40 20 Ml 0 0 100 C. Fig. S3 Comparison of leukaemia incidence rate in the triple targeted chimaeric mice and germline-transmission translocator mice

More information

CHAPTER 3 DEVELOPMENT OF DENV GROUP SPECIFIC REAL TIME RT-PCR

CHAPTER 3 DEVELOPMENT OF DENV GROUP SPECIFIC REAL TIME RT-PCR CHAPTER 3 DEVELOPMENT OF DENV GROUP SPECIFIC REAL TIME RT-PCR 28 Dengue is diagnosed by either detecting virus or antibody to the virus in blood. Isolation of virus in cell culture or in infant mouse brain

More information

WHEAT VARIETY GUIDE AVERY BRAWL CL PLUS BYRD DENALI HATCHER LANGIN SNOWMASS SUNSHINE THUNDER CL ANTERO

WHEAT VARIETY GUIDE AVERY BRAWL CL PLUS BYRD DENALI HATCHER LANGIN SNOWMASS SUNSHINE THUNDER CL ANTERO 2017 WHEAT VARIETY GUIDE AVERY BRAWL CL PLUS BYRD DENALI HATCHER LANGIN SNOWMASS SUNSHINE THUNDER CL ANTERO CONTENT GENETICS YOU CAN TRUST 1 COLORADO STATE UNIVERSITY WHEAT BREEDING & GENETICS PROGRAM

More information

Non-Organic-Based Isolation of Mammalian microrna using Norgen s microrna Purification Kit

Non-Organic-Based Isolation of Mammalian microrna using Norgen s microrna Purification Kit Application Note 13 RNA Sample Preparation Non-Organic-Based Isolation of Mammalian microrna using Norgen s microrna Purification Kit B. Lam, PhD 1, P. Roberts, MSc 1 Y. Haj-Ahmad, M.Sc., Ph.D 1,2 1 Norgen

More information

Roche Molecular Biochemicals Technical Note No. LC 10/2000

Roche Molecular Biochemicals Technical Note No. LC 10/2000 Roche Molecular Biochemicals Technical Note No. LC 10/2000 LightCycler Overview of LightCycler Quantification Methods 1. General Introduction Introduction Content Definitions This Technical Note will introduce

More information

PrimeScript RT reagent Kit (Perfect Real Time)

PrimeScript RT reagent Kit (Perfect Real Time) Cat. # RR037A For Research Use PrimeScript RT reagent Kit (Perfect Real Time) Product Manual Table of Contents I. Description... 3 II. Components... 3 III. Storage... 3 IV. Features... 4 V. Precautions...

More information

Cheatgrass Biology, Ecology, and Management

Cheatgrass Biology, Ecology, and Management Cheatgrass Biology, Ecology, and Management Fabián Menalled menalled@montana.edu 406-994-4783 Montana State http://ipm.montana.edu/cropweeds Presentation Outline Biology and identification of brome species

More information

Introduction To Real-Time Quantitative PCR (qpcr)

Introduction To Real-Time Quantitative PCR (qpcr) Introduction To Real-Time Quantitative PCR (qpcr) Samuel Rulli, Ph.D. Samuel.Rulli@QIAGEN.com Technical Support: BRCsupport@qiagen.com The products described in this webinar are intended for molecular

More information

PrimeScript 1st strand cdna Synthesis Kit

PrimeScript 1st strand cdna Synthesis Kit Cat. # 6110A For Research Use PrimeScript 1st strand cdna Synthesis Kit Product Manual Table of Contents I. Description... 3 II. Components... 3 III. Materials Required but not Provided... 3 IV. Storage...

More information

ON FARM UTILIZATION OF SOLID MANURE:DIFFERENCES IN

ON FARM UTILIZATION OF SOLID MANURE:DIFFERENCES IN ON FARM UTILIZATION OF SOLID MANURE:DIFFERENCES IN PLANT SPECIES NUMBER AND DRY MATER YIELD OF SEMI-NATURAL GRASSLAND FERTILIZED WITH MINERAL FERTILIZER AND SOLID MANURE Faculty of Agriculture, UNIVERSITY

More information

PrimeScript RT Master Mix (Perfect Real Time)

PrimeScript RT Master Mix (Perfect Real Time) Cat. # RR036A For Research Use PrimeScript RT Master Mix (Perfect Real Time) Product Manual Table of Contents I. Description... 3 II. Kit Components... 3 III. Materials Required but not Provided... 3 IV.

More information

Low cost and non-toxic genomic DNA extraction for use in molecular marker studies.

Low cost and non-toxic genomic DNA extraction for use in molecular marker studies. Low cost and non-toxic genomic DNA extraction for use in molecular marker studies. Version 1.4, February 28 th, 2013. Prepared by Bernhard Hofinger, Owen Huynh and Brad Till. 1. OBJECTIVE To develop and

More information

Absolute Human Telomere Length Quantification qpcr Assay Kit (AHTLQ) Catalog # reactions

Absolute Human Telomere Length Quantification qpcr Assay Kit (AHTLQ) Catalog # reactions Absolute Human Telomere Length Quantification qpcr Assay Kit (AHTLQ) Catalog #8918 100 reactions Product Description Telomeres are repetitive nucleotide elements at the ends of chromosomes that protect

More information

RUSSIAN WHEAT APHID USE THE GRDC FITE STRATEGY WHERE IS IT NOW?

RUSSIAN WHEAT APHID USE THE GRDC FITE STRATEGY WHERE IS IT NOW? RUSSIAN WHEAT APHID Russian wheat aphid (Diuraphis noxia) is a new pest to Australia, first detected in cereal crops in South Australia in May 2016. While grain growers are used to dealing with aphids

More information

Molecular Cell Biology - Problem Drill 11: Recombinant DNA

Molecular Cell Biology - Problem Drill 11: Recombinant DNA Molecular Cell Biology - Problem Drill 11: Recombinant DNA Question No. 1 of 10 1. Which of the following statements about the sources of DNA used for molecular cloning is correct? Question #1 (A) cdna

More information

Quantscript RT Kit. For first-strand cdna synthesis and two step RT-PCR.

Quantscript RT Kit. For first-strand cdna synthesis and two step RT-PCR. 1. Quantscript RT Kit For first-strand cdna synthesis and two step RT-PCR www.tiangen.com ER121221 Quantscript RT Kit Cat. no. KR103 Kit Contents Contents KR103-03 25 rxn KR103-04 100 rxn Quant Reverse

More information

TQ RXN 2X Q-PCR Master Mix (SYBR, no ROX) 1 ml x 2 TQ RXN 2X Q-PCR Master Mix (SYBR, no ROX) 1 ml x 5

TQ RXN 2X Q-PCR Master Mix (SYBR, no ROX) 1 ml x 2 TQ RXN 2X Q-PCR Master Mix (SYBR, no ROX) 1 ml x 5 www.smobio.com Product Information ExcelTaq series 2X Q-PCR Master Mix (SYBR, no ROX) TQ1100 200 RXN 2X Q-PCR Master Mix (SYBR, no ROX) 1 ml x 2 TQ1101 500 RXN 2X Q-PCR Master Mix (SYBR, no ROX) 1 ml x

More information

Bacteriophage MS2. genesig Standard Kit. Phage MS2 genome. 150 tests. Primerdesign Ltd. For general laboratory and research use only

Bacteriophage MS2. genesig Standard Kit. Phage MS2 genome. 150 tests. Primerdesign Ltd. For general laboratory and research use only TM Primerdesign Ltd Bacteriophage MS2 Phage MS2 genome genesig Standard Kit 150 tests For general laboratory and research use only 1 Introduction to Bacteriophage MS2 Bacteriophage MS2 is a non-enveloped,

More information

Functional Genomics Research Stream. Research Meeting: June 19, 2012 SYBR Green qpcr, Research Update

Functional Genomics Research Stream. Research Meeting: June 19, 2012 SYBR Green qpcr, Research Update Functional Genomics Research Stream Research Meeting: June 19, 2012 SYBR Green qpcr, Research Update Updates Alternate Lab Meeting Fridays 11:30-1:00 WEL 4.224 Welcome to attend either one Lab Log thanks

More information

HiPer Random Amplification of Polymorphic DNA (RAPD) Teaching Kit

HiPer Random Amplification of Polymorphic DNA (RAPD) Teaching Kit HiPer Random Amplification of Polymorphic DNA (RAPD) Teaching Kit Product Code: HTBM031 Number of experiments that can be performed: 5 Duration of Experiment: Protocol: 3.5 hours Agarose Gel Electrophoresis:

More information

Guide-it sgrna In Vitro Transcription and Screening Systems User Manual

Guide-it sgrna In Vitro Transcription and Screening Systems User Manual Clontech Laboratories, Inc. Guide-it sgrna In Vitro Transcription and Screening Systems User Manual Cat. Nos. 631438, 631439 & 631440 (042114) Clontech Laboratories, Inc. A Takara Bio Company 1290 Terra

More information

USB HotStart-IT. for increased specificity and consistent results. PCR, qpcr and qrt-pcr

USB HotStart-IT. for increased specificity and consistent results. PCR, qpcr and qrt-pcr USB HotStart-IT for increased specificity and consistent results PCR, qpcr and qrt-pcr USB PCR Reagents Choose USB HotStart-IT products for increased specificity and consistent results. Long and Accurate

More information

QuickExtract RNA Extraction Kit

QuickExtract RNA Extraction Kit Cat. Nos. QER09015 and QER090150 Connect with Epicentre on our blog (epicentral.blogspot.com), Facebook (facebook.com/epicentrebio), and Twitter (@EpicentreBio). www.epicentre.com Lit. # 299 12/2012 1

More information

Lecture Four. Molecular Approaches I: Nucleic Acids

Lecture Four. Molecular Approaches I: Nucleic Acids Lecture Four. Molecular Approaches I: Nucleic Acids I. Recombinant DNA and Gene Cloning Recombinant DNA is DNA that has been created artificially. DNA from two or more sources is incorporated into a single

More information

Recovery Plan for Wheat Blast (caused by Magnaporthe oryzae Triticum pathotype)

Recovery Plan for Wheat Blast (caused by Magnaporthe oryzae Triticum pathotype) Recovery Plan for Wheat Blast (caused by Magnaporthe oryzae Triticum pathotype) Contributors: William Bockus, Christian Cruz, Erick De Wolf, Jim Stack and Barbara Valent* (Kansas State University) Gary

More information

Roche Molecular Biochemicals Technical Note No. LC 9/2000

Roche Molecular Biochemicals Technical Note No. LC 9/2000 Roche Molecular Biochemicals Technical Note No. LC 9/2000 LightCycler Optimization Strategy Introduction Purpose of this Note Table of Contents The LightCycler system provides different detection formats

More information

Premix Ex Taq (Probe qpcr)

Premix Ex Taq (Probe qpcr) For Research Use Premix Ex Taq (Probe qpcr) Product Manual Table of Contents I. Description... 3 II. Principle... 4 III. Components... 5 IV. Materials Required but not Provided... 5 V. Storage... 5 VI.

More information

Introduction. Technical Note

Introduction. Technical Note DNA and RNA quantification: fast and simple with PicoGreen dsdna and RiboGreen RNA quantification reagents Fluorescence intensity on Infinite F2 and Infinite M2 Introduction DNA quantification Detection

More information

Kansas State University. Hays, KS USA

Kansas State University. Hays, KS USA Importance of Plant Resistance to Insect and Mite Vectors in Controlling Virus Diseases of Plants: Resistance to the Wheat Curl Mite (Acari: Eriophyidae)l, 2 T, L. Harvey, T. J. Martin, and D. L. Seifers

More information

Klamath Experiment Station

Klamath Experiment Station Cool-Season Grass Agroecozone Trial R.L. Dovel and J. Rainey' I ntroduction Irrigated pastures occupy over 95,000 acres in Klamath County and provide summer grazing for over 100,000 cattle. The currently

More information

Human Rhinovirus all subtypes (generic)

Human Rhinovirus all subtypes (generic) TM Primerdesign Ltd Human Rhinovirus all subtypes (generic) 5 non coding region (5 NCR) genesig Standard Kit 150 tests For general laboratory and research use only 1 Introduction to Human Rhinovirus all

More information

SuperPrep Cell Lysis & RT Kit for qpcr

SuperPrep Cell Lysis & RT Kit for qpcr SuperPrep Cell Lysis & RT Kit for qpcr 1402 F1267K SuperPrep Cell Lysis & RT Kit for qpcr SCQ-101 100 reactions Store at -20 C SuperPrep Cell Lysis Kit for qpcr SCQ-201 100 preparations Store at -20 C

More information

Cloning Small RNAs for Sequencing with 454 Technology

Cloning Small RNAs for Sequencing with 454 Technology Cloning Small RNAs for Sequencing with 454 Technology Protocol provided by Dr. Greg Hannon, Cold Spring Harbor Laboratory 1. RNA preparation 1. Total RNA is isolated from tissue or cells with TRIZOL followed

More information

TruSeq ChIP Sample Preparation

TruSeq ChIP Sample Preparation FOR RESEARCH USE ONLY Date: Illumina Kit Description: NOTE Unless familiar with the protocol in the latest version of the TruSeq ChIP Sample Preparation Guide (part # 15023092), new or less experienced

More information

Description...1 Components...1 Storage... 1 Technical Information...1 Protocol...2 Examples using the kit...4 Troubleshooting...

Description...1 Components...1 Storage... 1 Technical Information...1 Protocol...2 Examples using the kit...4 Troubleshooting... QuickClean II Gel Extraction Kit Cat. No. L00418 Technical Manual No. TM0594 Version: 03042011 I II III IV V VI VII VIII Description......1 Components.....1 Storage.... 1 Technical Information....1 Protocol.....2

More information

TaKaRa MiniBEST Viral RNA/DNA Extraction Kit Ver.5.0

TaKaRa MiniBEST Viral RNA/DNA Extraction Kit Ver.5.0 Cat. # 9766 For Research Use TaKaRa MiniBEST Viral RNA/DNA Extraction Kit Ver.5.0 Product Manual Table of Contents I. Description... 3 II. Components... 3 III. Storage and shipping... 3 IV. Preparation

More information

OsHV-1 detection and quantification by Real Time Polymerase Chain Reaction

OsHV-1 detection and quantification by Real Time Polymerase Chain Reaction European Union Reference Laboratory for Molluscs Diseases OsHV-1 detection and quantification by Real Time Polymerase Chain Reaction CONTENTS 1. SCOPE...2 2. REFERENCES...2 3. EQUIPMENT AND ENVIRONMENTAL

More information

SYBR Real-Time PCR Kit

SYBR Real-Time PCR Kit SYBR Real-Time PCR Kit For Amplification and detection of DNA in Quantitative real-time PCR (qpcr). Catalog No. QPG-040/QPG-041/QPG-042/QPG-043 User Manual Table of Contents Kit Contents and Storage...

More information

PureLink Plant Total DNA Purification Kit

PureLink Plant Total DNA Purification Kit USER GUIDE PureLink Plant Total DNA Purification Kit For purification of DNA from plant Catalog Number K1830-01 Document Part Number 25-0757 Publication Number MAN0000478 Revision 2.0 For Research Use

More information

Chromatrap 96 DNA Micro Elution Purification Kit

Chromatrap 96 DNA Micro Elution Purification Kit V 1.0 Chromatrap 96 DNA Micro Elution Purification Kit High throughput plates for ultra-pure DNA Protocol v1.0 Catalogue no. 500240 ADVANCEMENTS IN EPIGENETICS Contents Kit Components and Storage 3 Additional

More information

TECHNICAL BULLETIN. GenElute mrna Miniprep Kit. Catalog MRN 10 MRN 70

TECHNICAL BULLETIN. GenElute mrna Miniprep Kit. Catalog MRN 10 MRN 70 GenElute mrna Miniprep Kit Catalog Numbers MRN 10, MRN 70 TECHNICAL BULLETIN Product Description The GenElute mrna Miniprep Kit provides a simple and convenient way to purify polyadenylated mrna from previously

More information

FORAGE FERTILIZATION IN NORTH CAROLINA: CONCEPTS AND GUIDELINES

FORAGE FERTILIZATION IN NORTH CAROLINA: CONCEPTS AND GUIDELINES FORAGE FERTILIZATION IN NORTH CAROLINA: CONCEPTS AND GUIDELINES ESSENTIAL NUTRIENTS AND THE PRINCIPLE OF THE LIMITING FACTOR Forages, as all other plants, require seventeen elements for normal growth and

More information

QUANTITATIVE RT-PCR PROTOCOL (SYBR Green I) (Last Revised: April, 2007)

QUANTITATIVE RT-PCR PROTOCOL (SYBR Green I) (Last Revised: April, 2007) QUANTITATIVE RT-PCR PROTOCOL (SYBR Green I) (Last Revised: April, 007) Please contact Center for Plant Genomics (CPG) facility manager Hailing Jin (hljin@iastate.edu) regarding questions or corrections.

More information

Detection of Tomato yellow leaf curl virus Isolates by Multiplex. Polymerase Chain Reaction

Detection of Tomato yellow leaf curl virus Isolates by Multiplex. Polymerase Chain Reaction Technical Sheet No. 32 Detection of Tomato yellow leaf curl virus Isolates by Multiplex Polymerase Chain Reaction GENERAL Virus Detected: TYLCV isolates from tomato plants. DEVELOPED BY Name of researchers

More information

Analysis of the expression of homoeologous genes in polyploid cereals using fluorescence cdna-sscp

Analysis of the expression of homoeologous genes in polyploid cereals using fluorescence cdna-sscp Analysis of the expression of homoeologous genes in polyploid cereals using fluorescence cdna-sscp De Bustos A., Perez R., Jouve N. in Molina-Cano J.L. (ed.), Christou P. (ed.), Graner A. (ed.), Hammer

More information

Russian Wheat Aphid in Utah

Russian Wheat Aphid in Utah Page 1 of 5 Russian Wheat Aphid in Utah Fact Sheet No. 67 February 1989 Dr. Jay B Karren, Extension Entomologist Thomas A. Reeve, Extension Agent Introduction One of the most recent and important pests

More information

Israeli Acute Paralysis Virus

Israeli Acute Paralysis Virus TM Primerdesign Ltd Israeli Acute Paralysis Virus Helicase gene genesig Standard Kit 150 tests For general laboratory and research use only 1 Introduction to Israeli Acute Paralysis Virus Israeli acute

More information

Human T-lymphotropic Virus 1

Human T-lymphotropic Virus 1 TM Primerdesign Ltd Human T-lymphotropic Virus 1 Polymerase gene (POL) genesig Standard Kit 150 tests For general laboratory and research use only 1 Introduction to Human T-lymphotropic Virus 1 Human T-lymphotropic

More information

TOSCANA VIRUS oligomix Alert kit reagent for cdna "nested" amplification

TOSCANA VIRUS oligomix Alert kit reagent for cdna nested amplification TABLE OF CONTENTS INTENDED USE page 1 KIT DESCRIPTION page 1 KIT CHARACTERISTICS page 2 OTHER PRODUCTS REQUIRED page 2 MATERIALS PROVIDED IN THE KIT page 2 MATERIALS REQUIRED BUT NOT PROVIDED IN THE KIT

More information

Zika Virus. genesig Advanced Kit. Polyprotein gene. 150 tests. Primerdesign Ltd. For general laboratory and research use only

Zika Virus. genesig Advanced Kit. Polyprotein gene. 150 tests. Primerdesign Ltd. For general laboratory and research use only TM Primerdesign Ltd Zika Virus Polyprotein gene genesig Advanced Kit 150 tests For general laboratory and research use only 1 Introduction to Zika Virus Zika virus (ZIKV) is a member of the Flaviviridae

More information

BIOLOGY Dr.Locke Lecture# 27 An Introduction to Polymerase Chain Reaction (PCR)

BIOLOGY Dr.Locke Lecture# 27 An Introduction to Polymerase Chain Reaction (PCR) BIOLOGY 207 - Dr.Locke Lecture# 27 An Introduction to Polymerase Chain Reaction (PCR) Required readings and problems: Reading: Open Genetics, Chapter 8.1 Problems: Chapter 8 Optional Griffiths (2008) 9

More information

Human T-lymphotropic Virus 1

Human T-lymphotropic Virus 1 PCRmax Ltd TM qpcr test Human T-lymphotropic Virus 1 Polymerase gene (POL) 150 tests For general laboratory and research use only 1 Introduction to Human T-lymphotropic Virus 1 Human T-lymphotropic Virus

More information

GeneChip Eukaryotic Small Sample Target Labeling Assay Version II *

GeneChip Eukaryotic Small Sample Target Labeling Assay Version II * GENE EXPRESSION MONITORING TECHNICAL NOTE GeneChip Eukaryotic Small Sample Target Labeling Assay Version II * Introduction There is an overwhelming and continuing demand for a well characterized protocol

More information

Score winning cdna yields with SuperScript III RT

Score winning cdna yields with SuperScript III RT Score winning cdna yields with RT SuperScript III offers: Higher cdna yields Higher thermal stability Longer half-life than any other RT you could use Announcing SuperScript III Reverse Transcriptase How

More information

BioBank cdna kit. Instructions for the use of BioBank control cdna in real-time PCR

BioBank cdna kit. Instructions for the use of BioBank control cdna in real-time PCR BioBank cdna kit Instructions for the use of BioBank control cdna in real-time PCR Contents Introduction 3 Kit contents 4 Reagents and equipment to be supplied by user 4 Kit storage and stability 4 Primerdesign

More information

PRODUCT INFORMATION Thermo Scientific Luminaris Color Probe qpcr Master Mix #K0354 For 5000 rxns Lot Exp. Store at -20 C in the dark CERTIFICATE OF ANALYSIS The absence of endo-, exodeoxyribonucleases

More information

User Manual. NGS Library qpcr Quantification Kit (Illumina compatible)

User Manual. NGS Library qpcr Quantification Kit (Illumina compatible) NGS Library qpcr Quantification Kit (Illumina compatible) User Manual 384 Oyster Point Blvd, Suite 15 South San Francisco, CA 94080 Phone: 1 (888) MCLAB-88 Fax: 1 (650) 872-0253 www.mclab.com Contents

More information

Recombinant DNA Technology. The Role of Recombinant DNA Technology in Biotechnology. yeast. Biotechnology. Recombinant DNA technology.

Recombinant DNA Technology. The Role of Recombinant DNA Technology in Biotechnology. yeast. Biotechnology. Recombinant DNA technology. PowerPoint Lecture Presentations prepared by Mindy Miller-Kittrell, North Carolina State University C H A P T E R 8 Recombinant DNA Technology The Role of Recombinant DNA Technology in Biotechnology Biotechnology?

More information

User Manual. Chromofy dsdna binding dye Version 1.0 April 2009 For use in quantitative real-time PCR

User Manual. Chromofy dsdna binding dye Version 1.0 April 2009 For use in quantitative real-time PCR User Manual Chromofy dsdna binding dye Version 1.0 April 2009 For use in quantitative real-time PCR Chromofy dsdna binding dye Table of contents Background 4 Contents 4 Storage 4 Fluorescence data 4 Additionally

More information

The Polymerase Chain Reaction. Chapter 6: Background

The Polymerase Chain Reaction. Chapter 6: Background The Polymerase Chain Reaction Chapter 6: Background Invention of PCR Kary Mullis Mile marker 46.58 in April of 1983 Pulled off the road and outlined a way to conduct DNA replication in a tube Worked for

More information

Primerdesign TM Ltd. Dengue, Chikungunya, Zika Virus. (Multiplex kit) genesig kit. 100 tests. For general laboratory and research use only

Primerdesign TM Ltd. Dengue, Chikungunya, Zika Virus. (Multiplex kit) genesig kit. 100 tests. For general laboratory and research use only Primerdesign TM Ltd Dengue, Chikungunya, Zika Virus (Multiplex kit) genesig kit 100 tests For general laboratory and research use only Introduction to Flaviviruses Flavivirus is a genus of viruses in the

More information

Bauer Core Standard Protocol Title: Guidelines for Designing Real Time PCR Experiments Pages: 5 Revision: 1.1 Date: 4/15/04

Bauer Core Standard Protocol Title: Guidelines for Designing Real Time PCR Experiments Pages: 5 Revision: 1.1 Date: 4/15/04 Bauer Core Standard Protocol Title: Guidelines for Designing Real Time PCR Experiments Pages: 5 Revision: 1.1 Date: 4/15/04 Author(s): Claire Reardon Reviewers: Christian Daly Contact: claire@cgr.harvard.edu

More information

Sin Nombre Virus RT-PCR Detection Kit Product #51900

Sin Nombre Virus RT-PCR Detection Kit Product #51900 3430 Schmon Parkway Thorold, ON, Canada L2V 4Y6 Phone: 866-667-4362 (905) 227-8848 Fax: (905) 227-1061 Email: techsupport@norgenbiotek.com Sin Nombre Virus RT-PCR Detection Kit Product #51900 Product Insert

More information

10 RXN 50 RXN 500 RXN

10 RXN 50 RXN 500 RXN SeqPlex Enhanced DNA Amplification Kit for use with high throughput sequencing technologies Catalog Number SEQXE Storage Temperature 20 C TECHNICAL BULLETIN Product Description The SeqPlex DNA Amplification

More information

qpcr Quantitative PCR or Real-time PCR Gives a measurement of PCR product at end of each cycle real time

qpcr Quantitative PCR or Real-time PCR Gives a measurement of PCR product at end of each cycle real time qpcr qpcr Quantitative PCR or Real-time PCR Gives a measurement of PCR product at end of each cycle real time Differs from endpoint PCR gel on last cycle Used to determines relative amount of template

More information

TECHNICAL SHEET No. 18. Virus Detection: Potato Virus X (PVX)

TECHNICAL SHEET No. 18. Virus Detection: Potato Virus X (PVX) General Virus detected: PVX from potato leaves. General method: RT-PCR, PCR-ELISA. TECHNICAL SHEET No. 18 Virus Detection: Potato Virus X (PVX) Method: RT-PCR and PCR-ELISA Developed by Name of researchers:

More information

SYBR Premix DimerEraser (Perfect Real Time)

SYBR Premix DimerEraser (Perfect Real Time) Cat. # RR091A or Research Use SYBR Premix DimerEraser (Perfect Real Time) Product Manual Table of Contents I. Description... 3 II. Principle... 3 III. Components... 4 IV. Storage... 5 V. eatures... 5 VI.

More information