Nucleic Acid Purification

Transcription

1 Nucleic Acid Purification

2 Purification of Genomic DNA MasterPure Complete DNA and RNA Purification Kit Purify TNA, DNA, or RNA from any source The MasterPure Complete DNA and RNA Purification Kit enables efficient purification of intact total nucleic acid (TNA), DNA, or RNA from every type of biological material. The purified DNA is suitable for sequencing and other molecular biology applications. Purify DNA in 30 minutes without spin columns. A 260 /A 280 ratios consistently between 1.8 and 2.0. Recover >90% of theoretical DNA yield, including both low- and high-molecularweight nucleic acids. No phenol, chloroform, or other caustic solvents. Table 1. Typical yields obtained using the MasterPure Complete Kit. Sample Sample Size TNA µg DNA µg RNA µg HeLa/HL60 cells 1 x 10 6 cells Tissues Liver 5 mg Brain 5 mg Heart 5 mg Kidney 5 mg Thymus 5 mg Blood 200 µl Buffy coat 300 µl Mouse tail 0.5 cm E. coli 3.5 x 10 6 cells S. mutans 1.5 ml 0.9 Yeast* 2.2 x 10 6 cells (S. cerevisiae) 1.1 x 10 7 cells *For extraction of yeast DNA, the MasterPure Yeast DNA Purification Kit is recommended. MasterPure Complete DNA and RNA Purification Kit MC /10 Purifications MC Purifications For isolating TNA, DNA, or RNA. Contents: Red Cell Lysis Solution, Tissue and Cell Lysis Solution, MPC Protein Precipitation Reagent, 2X T&C Lysis Solution, TE Buffer, RNase A, RNase-Free DNase I, Proteinase K, 1X DNase Buffer, RiboGuard RNase Inhibitor. Note: MC89010 contains reagents for 10 TNA, 10 DNA, or 5 RNA purifications and MC85200 contains reagents for 200 TNA, 200 DNA, or 100 RNA purifications. 2

3 Purification of Genomic DNA MasterPure DNA Purification Kit for Blood Version II Purify DNA from whole blood or buffy coat The MasterPure DNA Purification Kit for Blood Version II can be used to purify genomic DNA from whole blood or buffy coat. With the optimized protocol for DNA, up to 100 µg of DNA can be recovered from 600 µl of buffy coat in less than 30 minutes. The purified DNA can be used for sequencing, PCR, digestion with restriction enzymes, Southern blotting, or other molecular biology applications. Purify DNA in 30 minutes without spin columns. A 260 /A 280 ratios consistently between 1.8 and 2.0. Recover >90% of theoretical DNA yield, including both low- and high-molecularweight nucleic acids. No phenol, chloroform, or other caustic solvents. Scalable protocol for large or small samples. Table 1. Yields of DNA using the MasterPure DNA Purification Kit for Blood Version II. Starting Material Yield 600 µl Buffy Coat µg 200 µl Whole Blood 3-9 µg 0.6 ml Whole Blood 9-27 µg 0.8 ml Whole Blood µg 1 ml Whole Blood µg 2 ml Whole Blood µg 3 ml Whole Blood µg 4 ml Whole Blood µg 5 ml Whole Blood µg MasterPure DNA Purification Kit for Blood Version II MB For 40 ml of whole blood MB For 400 ml of whole blood Contents: Red Cell Lysis Solution, Tissue and Cell Lysis Solution, MPC Protein Precipitation Reagent, RNase A, TE Buffer. techhelp@epicentre.com (800)

4 Purification of Genomic DNA MasterPure Plant Leaf DNA Purification Kit Purify DNA from challenging plant-leaf samples The MasterPure Plant Leaf DNA Purification Kit is designed to purify DNA from 35 to 100 mg of fresh plant-leaf tissue. The purified DNA can be used for sequencing, microsatellite typing, PCR amplification, restriction digestion, Southern blotting, or other molecular biology applications. Plants tested include Arabidopsis, apple, fern, grape, maize, pine, quillwort, sugarcane, sunflower, and tomato. Designed for plants high in polyphenolics, like grapevine. Scalable protocol takes less than 1 hour. Yields high-molecular-weight DNA. Figure 1. Comparative yields of DNA isolated with the MasterPure Plant Leaf Kit and another supplier s kit. MasterPure Kit Supplier G HMW DNA M M A 10-µl aliquot of each DNA preparation was separated by electrophoresis on a 0.7% agarose gel; the gel was stained with SYBR Gold and viewed with a Dark Reader Transilluminator. Lanes M, 1-kb DNA ladder; lanes 1-4, Pinot noir grape leaf DNA purified with the MasterPure Kit; lanes 5-8, Pinot noir grape leaf DNA purified with Supplier G Kit. HMW DNA, high-molecular-weight DNA. MasterPure Plant Leaf DNA Purification Kit MPP92010 MPP92100 Contents: Plant DNA Extraction Solution, Cleanup Solution, TE Buffer. 10 Purifications 100 Purifications 4

5 MasterPure Yeast DNA Purification Kit Purify DNA at high yields from yeast Purification of Genomic DNA The MasterPure Yeast DNA Purification Kit enables efficient, high-yield purification of high-molecular-weight DNA from yeast and other fungi. The protocol involves nonenzymatic cell lysis at 65 C, followed by removal of protein by precipitation, and nucleic acid precipitation and resuspension. The protocol can be easily scaled for larger or smaller samples, including single yeast colonies. The recovered nucleic acid can be used directly in most applications, including sequencing, PCR amplification, restriction endonuclease digestion, Southern blotting, and genomic library preparation. Higher yields of yeast chromosomal DNA than with other kits (Fig. 1). Recover DNA from a wide variety of yeast species, including Candida, Saccharomyces, Pichia, and Schizosaccharomyces, and filamentous fungi such as Aspergillus and Penicillium. Purify high-molecular-weight DNA in less than 40 minutes. No bead-beating, columns, or phenol or other organic extractions. Figure 1. Comparative yields of DNA. MasterPure Kit Supplier Q Supplier F MasterPure Kit Supplier Q Supplier F C. albicans S. cerevisiae 0 1,000 2,000 3,000 4,000 5,000 6,000 7,000 8,000 DNA, nanograms DNA was quantitated specifically with Hoechst fluorescent dye 33258, which gives minimal fluorescence with RNA. The data represent the average DNA yields determined by fluorometry from two experiments with S. cerevisiae and C. albicans. The MasterPure Kit produced up to 17 times more DNA from C. albicans and 12 times more DNA from S. cerevisiae than other kits. MasterPure Yeast DNA Purification Kit MPY Purifications MPY Purifications Contents: Yeast Cell Lysis Solution, MPC Protein Precipitation Reagent, TE Buffer, RNase A. techhelp@epicentre.com (800)

6 Purification of Genomic DNA MasterPure Gram-Positive DNA Purification Kit Purify genomic DNA from Gram-positive bacteria The MasterPure Gram-Positive DNA Purification Kit provides all of the reagents needed to purify DNA from Gram-positive bacteria. The purified DNA is suitable for fosmid library construction, PCR, restriction digests, Southern blotting, and other molecular biology applications. The protocol can also be adapted for total RNA purification. Scalable method for large sample volumes. Yields high-quality, high-molecular-weight DNA. Lysozyme included; no separate purchase required. Tested on a variety of species. Can also be used with Gram-negative bacteria. Figure 1. Electrophoretic analysis of DNA purified using the MasterPure Gram-Positive DNA Purification Kit. M 1 DNA purified from B. subtilis (ATCC 6051) was separated on a 1% agarose gel and stained with SYBR Gold. Lane M, kilobase ladder; lane 1, 300 ng of B. subtilis DNA. 12 kb 1 kb MasterPure Gram-Positive DNA Purification Kit MGP04020 MGP04100 Contents: Gram-Positive Cell Lysis Solution, MPC Protein Precipitation Reagent, Ready-Lyse Lysozyme, Proteinase K, TE Buffer, RNase A. 20 Reactions 100 Reactions 6

7 Meta-G-Nome DNA Isolation Kit Purify metagenomic DNA from water or soil samples Purification of Genomic DNA The Meta-G-Nome DNA Isolation Kit is used to isolate DNA from unculturable or difficultto-culture microbial species present in environmental water or soil samples. The simple extraction procedure involves low-speed centrifugation prior to filtration, and lysis with Ready-Lyse Lysozyme and Proteinase K. The kit isolates high-molecular-weight DNA that is randomly sheared (with fragment sizes approximately 40 kb), does not require size selection, and can be used directly for end-repair and construction of fosmid libraries. The kit can also be used to prepare 16S rrna metagenomic libraries for phylogenetic classification or species-abundance studies. Isolated DNA is also suitable for PCR (Fig. 1) or sequencing. No bead-beating, agarose plugs, pulse-field gel electrophoresis, or size selection. No phenol/chloroform extractions or CTAB. Figure 1. PCR amplification of soil metagenomic DNA. M DNA was isolated following the kit protocol, and PCR was performed using 16S rrna primers using various dilutions of the template DNA. Lane M, Kilobase ladder; lanes 1 and 2, 1 µl and 5 µl of PCR products from 1:10 template dilution; lanes 3 and 4, 1 µl and 5 µl of PCR products from 1:100 template dilution; lane 5, 1 µl of PCR products from undiluted template; lane 6, 1:10 dilution of sample from lane 5. Meta-G-Nome DNA Isolation Kit MGN Purifications Contents: Extraction buffer, Filter Wash Buffer, Meta-Lysis Solution (2X), MPC Protein Precipitation Reagent, Proteinase K, Fosmid Control DNA, RNase A, Ready-Lyse Lysozyme Solution, TE Buffer, 0.45-µm cellulose acetate filter membranes, 1.2-µm cellulose acetate filter membranes. techhelp@epicentre.com (800)

8 Purification of Genomic DNA WaterMaster DNA Purification Kit Purify DNA from water samples The WaterMaster DNA Purification Kit provides all of the reagents needed to purify microbial DNA from any water source after passage through a suitable filter. The DNA is suitable for PCR, cloning, sequencing, and DNA microarray analysis. Purifies high-molecular-weight DNA. Gentle procedure with no toxic solvents or mechanical cell disruption. Final DNA 50X more concentrated than competing kits. Figure 1. Detection of E. coli in water samples by PCR. M Varying amounts of E. coli K12 were diluted in 100-ml distilled water samples, trapped on 0.45-micron filters and DNA was extracted using the WaterMaster Kit. Aliquots of the extracted DNA were used as templates for PCR, using primers for a eubacterial rrna gene and the FailSafe PCR System. The PCR products were analyzed by agarose gel electrophoresis. Lane M, 100-bp ladder; PCR products from DNA purified from the indicated amounts of E. coli cells: lane 1, 10 5 ; lane 2, 10 3 ; lane 3, 10 2 ; lane 4, 10 1 ; lane 5, negative control. The amplicon is 350 bp. WaterMaster DNA Purification Kit WM Purifications WM Purifications Contents: WaterMaster Lysis Solution, MPC Protein Precipitation Reagent, TE Buffer, Proteinase K, RNase A, Spin Columns, Spin Resin. 8

9 FosmidMAX DNA Purification Kit BACMAX DNA Purification Kit Purify high-quality fosmid or BAC DNA Purification of Cloned DNA The FosmidMAX and BACMAX DNA Purification Kits were developed for easy, reliable isolation of high-quality fosmid or BAC DNA for many applications, including sequencing, fingerprinting, PCR, and preparation of shotgun libraries. No columns or binding resins that reduce yields and shear the DNA. No toxic organic solvents. Very low chromosomal DNA background. Ideal for isolating DNA from single-copy clones, and those induced using CopyControl Vectors. Figure 1. Quality of DNA prepared with the FosmidMAX Kit. Figure 2. Quality of DNA prepared with the BACMAX Kit. M M 1 10 Fosmid DNA isolated from six different fosmid clones (lanes 1 to 6) at single-copy number was digested with Hind III and analyzed by agarose gel electrophoresis. Lane M, Kilobase ladder. BAC DNA isolated from 10 different clones (lanes 1 to 10) was digested with Hind III and analyzed by agarose gel electrophoresis. Lane M, Kilobase ladder. FosmidMAX DNA Purification Kit FMAX046 Reagents sufficient for 150 x 1.5 ml; 10 x 40 ml; or 5 x 100 ml purifications. Contents: FosmidMAX Solutions 1-4, RiboShredder RNase Blend, TE Buffer. BACMAX DNA Purification Kit BMAX044 Reagents sufficient for 150 x 1.5 ml; 30 x 10 ml; 10 x 40 ml; or 5 x 100 ml purifications. Contents: BACMAX Solutions 1-4, RiboShredder RNase Blend, TE Buffer. 1 Kit 1 Kit techhelp@epicentre.com (800)

10 Purification of Cloned DNA BACMAX96 DNA Purification Kit Purify BAC or fosmid DNA in a 96-well format The BACMAX96 DNA Purification Kit provides easy, reliable isolation of high-quality BAC or fosmid DNA, that is free of contaminating chromosomal DNA, in a 96-well format. The DNA can be used for sequencing, fingerprinting, PCR, and the preparation of shotgun libraries. Isolate up to 1 µg of DNA from a single-copy BAC or fosmid clone and more than 2.5 µg from a CopyControl BAC clone. Purify 384 clones in less than 3 hours without automation. DNA is free of E. coli genomic DNA and RNA. Concentrated DNA obtained. Figure 1. Stability of autoinduced BAC and fosmid clones. A B DNA was isolated from seven different pcc1 BAC clones (A) and eight different pcc2 fosmid clones (B), from induced (+) and uninduced ( ) cultures, using the BACMAX96 Kit. For each preparation, an 8-μl aliquot was digested with Hind III and analyzed on a 1% agarose gel for 16 hours at 4 C. The identical fingerprints for induced and uninduced cultures show that autoinduction did not cause deletion or rearrangement. BACMAX96 DNA Purification Kit (with consumable plastics) BAC Kit Reagents sufficient for 4 x 96-well plates. Contents: BACMAX96 Solutions 1-3, BACMAX96 RNase Blend, Plamid-Safe ATP-dependent DNase, Plasmid-Safe 10X Buffer, 100 mm ATP Solution, TE Buffer, BAC and Fosmid Autoinduction Solutions, Bacterial Growth Plates, Filter Plates, Collection Plates, Gas Permeable Growth Plate Sealers, Standard Plate Sealers. 10

11 Direct Lysis Fosmid96 DNA Purification Kit Rapidly isolate fosmid DNA for sequencing Purification of Cloned DNA The Direct Lysis Fosmid96 Kit isolates fosmid DNA by direct lysis of a 200-µl overnight culture, eliminating the need for centrifugation of the culture and resuspension of the cell pellet. The protocol is complete in as little as 25 minutes. The kit is designed for rapid preparation of sequencing templates in a 96-well format from fosmid clones. No columns or binding resins that reduce yields and shear the DNA. No cell harvesting or resuspension of the cell pellet. Inoculation of culture in a 200-µl volume eliminates the need for 2-ml, deep-well growth plates. Reduces the amount of media used to <20 ml per plate. Figure 1. Restriction analysis of fosmid DNA. M 1 2 pcc1 Fosmid DNA 19 vector band Fosmid DNA was purified using the Direct Lysis Fosmid96 Kit and randomly chosen clones were digested with Not I. The DNA was analyzed by agarose gel electrophoresis. Lane M, Kilobase ladder; lane 1, 40-kb control insert; lanes 2-19, Not I-digested fosmid DNA. Direct Lysis Fosmid96 DNA Purification Kit FOS Kit Reagents sufficient for 5 x 96-well preps. Contents: Culture plates, 96-well fritted filter plates, collection plates, plate sealers, gas-permeable sealers, reagents for five 96-well preps. techhelp@epicentre.com (800)

12 Purification of Total RNA MasterPure RNA Purification Kit Purify RNA from any source The MasterPure RNA Purification Kit enables efficient purification of intact total RNA from a variety of biological materials. The process retains small RNAs that are typically lost when using column-based RNA purification. The purified RNA can be used directly in many molecular biology applications, e.g., RT-PCR, Northern blots, mirna analysis, or expression profiling with microarrays. No spin columns, phenol, chloroform, or other caustic solvents. Near-quantitative recovery of even small amounts of RNA. Table 1. RNA yields obtained using the MasterPure RNA Purification Kit. Sample Sample Size RNA Yield (µg) HeLa/HL60 cells 1 x 10 6 cells 7-15 Liver 5 mg Brain 5 mg 4-11 Heart 5 mg 4-5 Kidney 5 mg Thymus 5 mg 9-18 Buffy coat 300 µl 3-6 E. coli 3.5 x 10 6 cells Figure 1. Quality of RNA purified with the MasterPure RNA Purification Kit. M M 28 S 16 S Both adherent and suspension cell cultures were purified. A 200-ng aliquot of each RNA preparation was separated by denaturing formaldehyde agarose gel electrophoresis and visualized with SYBR Gold. Lane M, RNA Marker; lane 1, HeLa cell RNA; lane 2, Ramos cell RNA; lane 3, BDCM cell RNA; lane 4, NIH3T3 cell RNA; lane 5, NRK cell RNA. MasterPure RNA Purification Kit MCR Purifications Contents: Red Cell Lysis Solution, Tissue and Cell Lysis Solution, MPC Protein Precipitation Reagent, 2X T&C Lysis Solution, TE Buffer, RNase-Free DNase I, Proteinase K, 1X DNase Buffer, RiboGuard RNase Inhibitor. 12

13 MasterPure Yeast RNA Purification Kit Purify RNA from yeast and filamentous fungi Purification of Total RNA The MasterPure Yeast RNA Purification Kit provides all of the reagents needed to purify RNA from yeast cell types (including Candida, Saccharomyces, Schizosaccharomyces, Pichia) and filamentous fungi. The kit uses a rapid salting-out process to remove contaminating macromolecules. The purified RNA is suitable for cdna synthesis and microarray gene expression analysis. No hot acid phenol, spin columns, or bead-beating. Faster than spheroplasting. No extra enzymes or equipment to purchase. Yields µg RNA from 1 ml of mid-log S. cerevisiae. Figure 1. Purity of RNA. RNA was extracted from S. cerevisiae using the MasterPure Kit and stored at 20 C for 2 months before analysis on an Agilent Technologies 2100 Bioanalyzer. The electropherogram demonstrates the high-purity, intact RNA. MasterPure Yeast RNA Purification Kit MPY Reactions MPY Reactions Contents: Extraction Reagent for RNA, MPC Protein Precipitation Reagent, TE Buffer (in 100-reaction kit only), Proteinase K, RNase-Free DNase I, RiboGuard RNase Inhibitor, 10X DNase Buffer, 2X T & C Lysis Solution. techhelp@epicentre.com (800)

14 Purification of Total RNA MasterPure Plant RNA Purification Kit Purify high-quality RNA from plant tissues The MasterPure Plant RNA Purification Kit enables purification of intact total RNA from plant tissues. The RNA obtained is suitable for many applications, such as RNA-Seq, RT-PCR, Northern blots, mirna analysis, and expression profiling with microarrays. Unlike other plant RNA purification kits, a DNase I treatment step is included. Plants tested include banana, citrus, cotton, geranium, grape, maize, maple, raspberry, soybean, strawberry, and tomato leaves, alfalfa sprouts, soybean seedlings and seeds, maize seedlings and mature roots, pine needles, and field mustard seeds. Optimized technology for plant tissues to remove polyphenols and polysaccharides. No spin columns that reduce yields; small RNAs are purified effectively. Scalable protocol takes less than 1 hour. No phenol, chloroform, or other caustic solvents. Figure 1. Purification of intact plant RNA from a variety of plant tissues. M M Plant tissue RNA was purified using the MasterPure Plant RNA Purification Kit, and 500 ng of each type of RNA was separated on a 1% formaldehyde gel and visualized with SYBR Gold I. Lanes M, RNA ladder; lane 1, maize seedling RNA; lane 2, grape leaf RNA; lane 3, alfalfa sprout RNA; lane 4, strawberry leaf RNA; lane 5, raspberry leaf RNA; lane 6, pine needle RNA; lane 7, maize root RNA. MasterPure Plant RNA Purification Kit MPR Purifications MPR Purifications Contents: Plant Tissue and Cell Lysis Solution, 2X T and C Lysis Solution, MPC Protein Precipitation Reagent, RNase-Free Water, RNase-Free DNase 1, Proteinase K, RiboGuard RNase Inhibitor, 100 mm DTT, 10X DNase Buffer. 14

15 ExtractMaster Fecal DNA Extraction Kit Extract PCR-ready DNA from fecal samples Rapid Extraction of PCR-Ready DNA The ExtractMaster Fecal DNA Extraction Kit provides all of the reagents necessary to recover PCR-ready DNA from fecal samples. The kit removes small-molecule inhibitors that often coextract with DNA from such samples. The extracted DNA can be used to amplify bacterial, protist, or animal templates for a variety of physiological and pathophysiological research applications. Gentle procedure with no toxic solvents or mechanical cell disruption. Final DNA recovered in a small volume, making downstream analysis easier. Figure 1. PCR analysis of fecal DNA extracts. M Figure 2. Identification of rotavirus from infant fecal samples. M Extracted DNA was amplified using PCR using primers for a eubacterial rrna gene. Lane M, 100-bp ladder; lane1, human fecal DNA; lane 2, rat fecal DNA; lane 3, rooster fecal DNA; lane 4, no template. The amplicon is 350 bp. Rotavirus dsrna was isolated using the ExtractMaster Kit and used as templates for RT-PCR. Lane M, 100-bp ladder; lanes 1 and 3, RT-PCR products from nucleic acids isolated by phenol-chloroform extraction; lanes 2 and 4, RT-PCR products from nucleic acids isolated using the ExtractMaster Kit. The 1,062-bp amplicon that appears in lanes 2 and 4 corresponds to the full-length vp7 RNA, ExtractMaster Fecal DNA Extraction Kit FD Purifications FD Purifications Contents: Fecal DNA Extraction Buffer, Proteinase K, Fecal Lysis Buffer, Protein Precipitation Reagent, DNA Precipitation Solution, Pellet Wash Solution, TE Buffer, Inhibitor Removal Resin, Spin Columns. techhelp@epicentre.com (800)

16 Rapid Extraction of Nucleic Acids QuickExtract DNA Extraction Kits Rapidly prepare PCR-ready DNA from various sample types QuickExtract products offer a rapid and efficient method for extracting genomic DNA from virtually any sample for PCR-based assays. Most samples can be processed in 8 minutes with only two sequential heating steps. Fast PCR-ready DNA in 3-8 minutes, ideal for high-throughput workflows. Safe No organic extraction. Efficient No columns, transfers, or sample loss. Figure 1. FailSafe PCR amplification of extracted DNA M Buccal cells were extracted using the BuccalAmp DNA Extraction Kit, and all other samples with QuickExtract DNA Extraction Solution. PCR was performed using primers to amplify the regions indicated: Lanes 1-3, human β-globin (human buccal cells, HeLa cells, and human hair follicle, respectively); lane 4, transgenic mouse GAPDH (mouse tail snip); lane 5, E. coli 16S ribosomal RNA gene (bacteria); lane 6, transgenic SV40 T antigen (mouse tail snip). Figure 2. PCR amplification of FFPE DNA M M DNA was extracted from a slide-mounted, FFPEpreserved human skeletal muscle tissue section with the QuickExtract FFPE DNA Extraction Kit. Two microliters of undiluted, extracted DNA was amplified with primers for three different loci: tumor protein 53 (TP53), dystrophin (DMD), and tumor necrosis factor (TNF). The products were separated on a 3% agarose gel and were visualized with SYBR Gold. Lane M, 100-bp DNA ladder; lane 1, exon 2 of TP53; lane 2, exon 3 of TP53; lane 3, exon 11 of TP53; lane 4, exon 6 of DMD; lane 5, exon 50 of DMD; lane 6, exon 3 of DMD; lane 7, exon 4 of TNF. QuickExtract DNA Extraction Solution QE0905T QE ml 50 ml 16

17 Rapid Extraction of Nucleic Acids QuickExtract Family of Products Products Time Applications Samples Tested QuickExtract DNA Extraction Solution (QE0905T, QE09050) BuccalAmp DNA Extraction Kits (BQ0901SCR, BQ0908SCR, BQ0916SCR) QuickExtract RNA Extraction Kit (QER09015, QER090150) QuickExtract Plant DNA Extraction Solution (QEP80705, QEP70750) QuickExtract Seed DNA Extraction Solution (QES08095T, QES080950) QuickExtract Bacterial DNA Extraction Kit (QEB0905T, QEB09050) QuickExtract FFPE DNA Extraction Kit (QEF81805, QEF81050) QuickExtract FFPE RNA Extraction Kit (QFR82805, QFR82050) 3-8 min Genotyping, genetic studies, identity testing, viral/microbial screening 3-8 min Genotyping, genetic studies, identity testing Hair follicles, feathers (quillend cells), tissue-culture cells, buccal cells, zebrafish (organs, scales), mouse tail snips Buccal cells from human or animal subjects. Catch-All Buccal Swabs included. 6 min RT-PCR Cultured cells: human, mouse, rat, E. coli, S. aureus 8 min PCR, e.g., GMO testing Arabidopsis, barley, maize, emmer, pepper, rice, spelt, spinach, soybeans, wheat 8 min PCR, e.g., GMO testing Apple, cotton, sunflower, tomato, barley, maize, oats, rice, rye, wheat 15 min PCR, restriction digests, PFGE, optical mapping 62 min Microsatellite, single-nucleotide polymorphisms (SNP), tumor heterogeneity studies, copy number variations (CNV), methylation analysis, short tandem repeats (STR) Gram-positive: Bacillus subtilis Bifidobacterium spp Brevibacterium linens Clostridium perfringens Lactobacillious plantarum Listeria monocytogenes Staphylococcus equorum Streptococcus agalactiae Streptococcus pyrogenes Streptococcus thermophilus Gram-negative: E. coli Salmonella enterica Salmonella typhimurium Vibrio gazogenes Formalin-fixed, paraffinembedded human tissue samples 32 min RT-PCR Formalin-fixed, paraffinembedded human tissue samples For more information, see: techhelp@epicentre.com (800)

18 Rapid Extraction of PCR-Ready DNA BuccalAmp DNA Extraction Kit Extract PCR-ready DNA from buccal samples The BuccalAmp DNA Extraction Kit is a single-tube system for rapid preparation of DNA from human or other mammalian buccal (cheek) swab samples for PCR-based assays, such as human or animal identity testing, and SNP analysis. The kits incorporate the QuickExtract DNA Extraction Solution that permits processing of samples using a simplified protocol. The kits provide a choice of foam Catch-All Swabs or MasterAmp brushes (see next page), and include aliquots of the QuickExtract Solution as individual sample tubes. Gentle sample collection using a swab made of soft, porous foam on a flexible plastic handle; alternative to blood draws. Reduces risks from sample collection (e.g., needle sticks) and uses no phenol, chloroform, or other toxic solvents. Suitable for high-throughput and automated assays. Figure 1. BuccalAmp extraction protocol. Rotate swab on the inside of cheek Rotate swab in QuickExtract DNA Extraction Solution Heat at 65ºC for 1 minute and 98ºC for 2 minutes PCR-ready DNA Figure 2. PCR amplification of extracted DNA. M 1 18 DNA was extracted from 18 self-collected samples with the BuccalAmp DNA Extraction Kit, and amplified using primers to the β-globin gene. Lane M, 100-bp DNA ladder; lanes 1-18, representative PCR results. 18

19 Rapid Extraction of PCR-Ready DNA Buccal Swabs and Brushes Epicentre s buccal swabs and brushes are designed for safe, gentle buccal cell collection for subsequent DNA extraction and PCR analysis. Both swabs and brushes are provided individually packaged in sterile hard-pack plastic cylinders (hard pack) or as a soft pack. Catch-All Sample Collection Swabs are soft foam swabs on soft, flexible plastic handles. Catch-All Swabs provide gentle, safe buccal sample collection, even for infants, and the porous foam on these swabs catches more of the sample than buccal brushes. MasterAmp Buccal Brushes are soft bristle-type buccal brushes. BuccalAmp DNA Extraction Kits with Catch-All Hard Pack Buccal Swab BQ0901SCR 1 Kit (15 Tubes & Swabs) BQ0908SCR 8 Kits (120 Tubes & Swabs) BQ0916SCR 16 Kits (240 Tubes & Swabs) BuccalAmp DNA Extraction Kits with Catch-All Soft Pack Buccal Swab BQ0901SSC 1 Kit (15 Tubes & Swabs) BQ0908SSC 8 Kits (120 Tubes & Swabs) BQ0916SSC 16 Kits (240 Tubes & Swabs) BuccalAmp DNA Extraction Kits with MasterAmp Brush Hard Pack BQ0901SRB 1 Kit (15 Tubes & Brushes) BQ0908SRB 8 Kits (120 Tubes & Brushes) BQ0916SRB 16 Kits (240 Tubes & Brushes) BuccalAmp DNA Extraction Kits with MasterAmp Brush Soft Pack BQ0901SBS 1 Kit (15 Tubes & Brushes) BQ0908SBS 8 Kits (120 Tubes & Brushes) BQ0916SBS 16 Kits (240 Tubes & Brushes) Catch-All Sample Collection Swab (Hard Pack) QEC091H Catch-All Sample Collection Swab (Soft Pack) QEC89100 MasterAmp Buccal Brush (Hard Pack) MB100BR MasterAmp Buccal Brush (Soft Pack) MB100SP 100 buccal swabs in hard-pack plastic carrier 100 buccal swabs 100 buccal brushes in hard-pack plastic carrier 100 buccal brushes techhelp@epicentre.com (800)

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