Supporting Information
|
|
- Theresa Thornton
- 6 years ago
- Views:
Transcription
1 Supporting Information Wiley-VCH Weinheim, Germany
2 A new homogenous assay for studying mira maturation Brian Patrick Davies and Christoph Arenz General Information For MALDI-TF measurements a Voyager-DE Pro Biospectrometry Workstation from PerSeptive Biosystems (Foster City, USA) mass spectrometer was used in linear und negative ion mode. Reversephase HPLC was carried out with a Gilson semi-preparative HPLC fitted with an analytical Polaris column from Varian Inc. (5µm, 2Å, 25mm x 4.6mm). Fluorescence measurements were done on a Varian Cary Eclipse fluorescence spectrophotometer or a BMG Labtech Fluorostar ptima plate reader. ptical analyses were performed on a Varian Cary 1 Bio-UV/VIS spectrophotometer. Recombinant human Dicer was supplied by Stratagene or Gene Therapy Systems (Genlantis). Formation of the 72-mer pre-mira sequence by ligation of the labeled 5 - and 3 -terminal partial strands. The following oligonucleotides were purchased from IBA GmbH, Göttingen, Germany: 1) 5 -FAM-36mer (FAM-EX-5- GGCAAAUUGAGGUAGUAGGUUGUAUAGUAGUAAUUA) 1 and 2) 3 -DABCYL-36mer with 5 -phosphate 2 (phosphate-cacaucauacuauacaaugugcuagcuuucuuugcu-dabcyl) H CH H S H H CH 2 P 5'-ligo 3 C 6 - CH H 3'-ligo P - H H H Ligation of 5 - and 3 -terminal partial strands (T4 Ligase from ew England Biolabs): 1 nmol reaction: 5 -FAM-36mer 1 µl (.1 nmol/ µl) 3 -DABCYL-36mer 1µL (.1 nmol/ µl) T4 Ligase buffer (1x) 24 µl RAsin (4U/µL) 5 µl T4 RA ligase (2U/µL) 1 ml Total volume 239µL Reaction time: 18 hours at 37 C.
3 Analysis of ligation product: 2 % denaturing urea PAGE-gels as well as native PAGE-Gels in TBE buffer were used. Stain: SYBR GREE (Molecular Probes) a b c Figure S1: a) Denaturing PAGE from ligation, Column 1: both partial strands mixed, Column 2: reaction mixture after 18 hours; b) native PAGE without SYBR Green staining, Column 1: 5 -FAM-36mer, Column 2: 3 -DABCYL-36mer, Column 3: both partial strands mixed, Column 4: ligation mixture; c) ative PAGE stained with SYBR Green (1:1 dilution in TBE buffer), labeled as in b). Purification of ligation product: The ligation product was first purified by phenol extraction and aac precipitation in isopropanol. The raw product was further purified by reverse-phase HPLC and lyophilized. MALDI-Massenspektrometrie: Calculated: [M-H]/z 24411, [M-2H]/2z 1225, [M-3H]/3z Found: [M-H]/z 24432, [M-2H]/2z 1221, [M-3H]/3z Voyager Spec #1 MC=>MC[BP = , 957] % Intensity Mass (m/z) Figure S2: MALDI-TF spectrum of ligation product (pre-mira beacon).
4 In vitro Transcription A Promega T7 RiboMAX Express in vitro transcription kit was used. According to instructions the template containing the T7-primer was used. The DA template with underlined T7-promoter sequence is as follows: 5 -AGCAAAGAAAGCTAGCACATTGTATAGTATGATGTGTAATTACTACTATACAACCTACTACCTCAA TTTGCCTATAGTGAGTCGTATTA-3 The 5 -phosphates were then removed with alkaline phosphatase (ew England Biolabs) and rephosphorylated with polynucleotide kinase (ew England Biolabs). The final sequence was purified as described above using reverse-phase HPLC and analyzed by MALDI-TF. Fluorescence Assay Measurements on Varian Cary Eclipse fluorescence spectrophotometer: a) With human recombinant Dicer A total volume of 1mL contained 5nM RA beacon, 15mM acl, 2 mm Tris-HCl, 2.5mM MgCl 2, 1 mm DTT and either 25 U recombinant Dicer or heat denatured Dicer. The fluorescence was measured every 1 minutes over the course of 4-6 hours. b) With cell lysate from HEK 293 cells: A total volume of 1.1mL contained 14nM RA beacon, 15mM acl, 2mM Tris-HCl, 2.5mM MgCl 2 and either 2µL cell lysate or 2µL heat denatured cell lysate. The fluorescence was measured every 1 minutes over the course of 4 hours Figure S3: PAGE analysis after incubation with Dicer, Column 1: control with beacon, Column 2: beacon digest, Column 3: control with unlabeled pre-let7 transcript, Column 4: digest of unlabeled pre-let7 transcript. The black arrows show the mira bands (21-23mer), the green arrow shows the cleaved 5 -fluorescein end.
5 Incubation with potential inhibitors of mira maturation Measurement on BMG Labtech Fluorostar ptima plate reader: A total volume of 1µL contained 3nM RA-Sonde, 2 mm Tris-HCl, 2.5mM MgCl 2, 15mM acl, 1mM DTT, 1µM peptide/kanamycin and.5 U recombinant Dicer ormal Peptide S117 Peptide S186 Peptide S417 Kanamycin 2 F t /min Figure S4: Dicer cleavage without inhibitor or in the presence of 1µM peptide or kanamycin, without preincubation of RA and recombinant Dicer. The initial rise in fluorescence in the presence of peptide S186 can be avoided by incubating for 3 minutes with the RA before adding Dicer to the reaction. Sequences of the added peptides: Peptide S117: Ac-H-SSIYALEPDQKG-CH 2 Peptide S186: Ac-H-AKPYSQRRKTSG-CH 2 Peptide S417: Ac-H-RYIKKEFEFG-CH 2 Please note: The peptides correspond to the dicer sequence, but the - and C- termini are exchanged. Cell lysate from HEK293 cells Cells were taken up in buffer containing 2mM Tris-HCl ph 7.6, 75mM acl, 5mM MgCl 2, 2mM DTT, 1% Glycerol and Roche protease inhibitor and lyophilized by ultrasound. The mixture was centrifuged first for 1 minutes at 43 g and 4 C. The supernatant was then centrifuged for 5 minutes at 12,1 g and room temperature. The protein concentration in the supernatant was determined by the Bradford assay [1] (typically between 1 und 25 µg/ml). The cell lysate was stored by -2 C. The cell lysate for control measurements was heated at 95 C for 2 minutes and also stored at -2 C. References: [1] M. M. Bradford, Anal Biochem 1976, 72, 248.
Supporting Information
Supporting Information Wiley-VCH 2006 69451 Weinheim, Germany Rolling-circle Amplification of a DNA Nanojunction Chenxiang Lin, Mingyi Xie, Julian J.L. Chen, Yan Liu and Hao Yan A. RCA replication of the
More informationXactEdit Cas9 Nuclease with NLS User Manual
XactEdit Cas9 Nuclease with NLS User Manual An RNA-guided recombinant endonuclease for efficient targeted DNA cleavage Catalog Numbers CE1000-50K, CE1000-50, CE1000-250, CE1001-250, CE1001-1000 Table of
More informationElectronic Supplementary Information
Electronic Supplementary Information Ultrasensitive quantification of mature micrornas by real-time PCR based on ligation of ribonucleotide-modified DNA probe Jiangyan Zhang, Zhengping Li,* Hui Wang, Yucong
More informationSensoLyte 620 HCV Protease Assay Kit *Fluorimetric*
SensoLyte 620 HCV Protease Assay Kit *Fluorimetric* Catalog # 71146 Kit Size 100 Assays (96-well plate) Convenient Format: Complete kit including all the assay components. Optimized Performance: Optimal
More informationSUPPORTING INFORMATION. A cleavage-responsive stem-loop hairpin for assaying guide RNA activity
SUPPORTING INFORMATION A cleavage-responsive stem-loop hairpin for assaying guide RNA activity Tara R. deboer 1, Noreen Wauford 1, Jing-Yi Chung, Miguel Salvador Torres Perez, and Niren Murthy* University
More informationTorsional Constraints of DNA Substrates Impact Cas9 Cleavage
Supporting Information Torsional Constraints of DNA Substrates Impact Cas9 Cleavage Michael H. Räz, Kumi Hidaka, Shana J. Sturla, Hiroshi Sugiyama, *,, and Masayuki Endo *, Institute for Integrated Cell-Material
More informationAmpliScribe T7-Flash Transcription Kit
AmpliScribe T7-Flash Transcription Kit Cat. Nos. ASF3257 and ASF3507 Available exclusively thru Lucigen. lucigen.com/epibio www.lucigen.com MA191E AmpliScribe T7-Flash Transcription Kit 12/2016 1 1. Introduction
More informationDNA 5 End-Labeling System INSTRUCTIONS FOR USE OF PRODUCT U2010.
Technical Bulletin DNA 5 End-Labeling System INSTRUCTIONS FOR USE OF PRODUCT U2010. PRINTED IN USA. Revised 12/12 DNA 5 End-Labeling System All technical literature is available on the Internet at: www.promega.com/protocols/
More informationLigation Independent Cloning (LIC) Procedure
Ligation Independent Cloning (LIC) Procedure Ligation Independent Cloning (LIC) LIC cloning allows insertion of DNA fragments without using restriction enzymes into specific vectors containing engineered
More information3-Input Majority Logic Gate and Multiple Input Logic Circuit Based on DNA Strand Displacement
Supporting Information for 3-Input Majority Logic Gate and Multiple Input Logic Circuit Based on DNA Strand Displacement Wei Li, Yang Yang, Hao Yan, Yan Liu Department of Chemistry and Biochemistry and
More informationFisher (Fairlawn, NJ) and Sigma-Aldrich (St. Louis, MO) and were used without further. (Promega) and DpnI (New England Biolabs, Beverly, MA).
175 Appendix III Chapter 4 Methods General. Unless otherwise noted, reagents were purchased from the commercial suppliers Fisher (Fairlawn, NJ) and Sigma-Aldrich (St. Louis, MO) and were used without further
More informationSensoLyte 490 HCV Protease Assay Kit *Fluorimetric*
SensoLyte 490 HCV Protease Assay Kit *Fluorimetric* Catalog # 72087 Unit Size Kit Size 1 Kit 200 Assays (96-well) or 500 assays (384-well) This kit is optimized to detect the activity of Hepatitis C Virus
More informationBACTERIAL PRODUCTION EXPRESSION METHOD OVERVIEW: PEF # GENE NAME EXPRESSION VECTOR MOLECULAR WEIGHT kda (full-length) 34.
BACTERIAL PRODUCTION PEF # GENE NAME EXPRESSION VECTOR MOLECULAR WEIGHT 2015-XXXX XXXX pet-32a 50.9 kda (full-length) 34.0 kda (cleaved) EXPRESSION METHOD OVERVIEW: Plasmid DNA was transformed into BL21
More informationOne-Electron Oxidation of DNA: Thymine versus Guanine Reactivity
One-Electron Oxidation of DNA: Thymine versus Guanine Reactivity Sriram Kanvah and Gary B Schuster SUPPORTING INFORMATION EXPERIMENTAL PROCEDURES T4 polynucleotide Kinase (T4 PNK) was purchased from New
More informationComparison of different methods for purification analysis of a green fluorescent Strep-tag fusion protein. Application
Comparison of different methods for purification analysis of a green fluorescent Strep-tag fusion protein Application Petra Sebastian Meike Kuschel Stefan Schmidt Abstract This Application Note describes
More informationSequencing of DNA lesions facilitated by site-specific excision via base. excision repair DNA glycosylases yielding ligatable gaps
Supporting information Sequencing of DNA lesions facilitated by site-specific excision via base excision repair DNA glycosylases yielding ligatable gaps Jan Riedl, Aaron M. Fleming, and Cynthia J. Burrows*
More information+ M III. IMAP Screening Express Kit Product #8073 Quantity: 8000, 20 µl reactions (80 µl final volumes) Low FP. M III High FP.
Product Insert IMAP Screening Express Kit Product #8073 Quantity: 8000, 20 µl reactions (80 µl final volumes) Introduction About the IMAP Screening Express Kit The IMAP Screening Express Kit is used to
More informationcatalytic hairpin DNA assembly for dual-signal amplification toward homogenous analysis of protein and
Electronic Supplementary Material (ESI) for ChemComm. This journal is The Royal Society of Chemistry 2015 Supporting Information Programmable Mg 2+ -dependent DNAzyme switch by the catalytic hairpin DNA
More informationCircLigase ssdna Ligase
Cat. Nos. CL4111K and CL4115K Connect with Epicentre on our blog (epicentral.blogspot.com), Facebook (facebook.com/epicentrebio), and Twitter (@EpicentreBio). www.epicentre.com Lit. # 222 10/2012 1 EPILIT222
More informationDesigned thiazole orange nucleotides for the synthesis of single labelled oligonucleotides that fluorescence upon matched hybridization
(ESI) for Organic and Biomolecular Chemistry Designed thiazole orange nucleotides for the synthesis of single labelled oligonucleotides that fluorescence upon matched hybridization Lucas Bethge, a Ishwar
More informationZR-96 Genomic DNA Clean & Concentrator -5 Catalog Nos. D4066 & D4067
INSTRUCTION MANUAL ZR-96 Genomic DNA Clean & Concentrator -5 Catalog Nos. D4066 & D4067 Highlights 96-well plate recovery of large-sized DNA (e.g., genomic, mitochondrial, plasmid (BAC/PAC), viral, phage,
More informationLabeling Protocol for mytags Immortal Libraries
5840 Interface Drive, Suite 101 Ann Arbor MI 48103 1 (734) 998 0751 techsupport@arborbiosci.com Labeling Protocol for mytags Immortal Libraries March 2018 Version 1.5 Contents Reagents and Equipment...
More informationNEBNext RNase III RNA Fragmentation Module
SAMPLE PREPARATION NEBNext RNase III RNA Fragmentation Module Instruction Manual NEB #E6146S 100 reactions NEBNext RNase III RNA Fragmentation Module Table of Contents: Description....2 Applications....2
More informationSupplementary Figure S1 Supplementary Figure S2 Supplementary Figure S3. Supplementary Figure S4
Supplementary Figure S1 Supplementary Figure S2 Supplementary Figure S3 Supplementary Figure S4 Supplementary Figure S5 Supplementary Figure S6 Supplementary Figure S7 Supplementary Figure S8 Supplementary
More informationReverTra 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 informationViral RNAi suppressor reversibly binds sirna to. outcompete Dicer and RISC via multiple-turnover
Supplementary Data Viral RNAi suppressor reversibly binds sirna to outcompete Dicer and RISC via multiple-turnover Renata A. Rawlings 1,2, Vishalakshi Krishnan 2 and Nils G. Walter 2 * 1 Biophysics and
More informationCloning small RNAs for Solexa Sequencing version 2.0 by Nelson Lau Page 1 of 5 (Modified from Solexa sequencing protocol from Bartel lab)
Cloning small RNAs for Solexa Sequencing version 2.0 by Nelson Lau 09162008 Page 1 of 5 General Cloning Protocol: Gel-purification 1. Pour 1mm thick, urea denaturing 10% or 15% polyacrylamide gels, with
More informationFOR 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 informationSupporting Information
Supporting Information Development of a 2,4-Dinitrotoluene-Responsive Synthetic Riboswitch in E. coli cells Molly E. Davidson, Svetlana V. Harbaugh, Yaroslav G. Chushak, Morley O. Stone, Nancy Kelley-
More informationSupporting Information
Supporting Information Efficient RNA synthesis by in vitro transcription of a triazole-modified DNA template Afaf H. El-Sagheer a,b and Tom Brown a a School of Chemistry, University of Southampton, Highfield,
More informationCircLigase II ssdna Ligase
Cat. Nos. CL9021K and CL9025K Connect with Epicentre on our blog (epicentral.blogspot.com), Facebook (facebook.com/epicentrebio), and Twitter (@EpicentreBio). www.epicentre.com Lit. # 298 12/2012 1 EPILIT298
More informationSupplementary Information: Materials and Methods. Immunoblot and immunoprecipitation. Cells were washed in phosphate buffered
Supplementary Information: Materials and Methods Immunoblot and immunoprecipitation. Cells were washed in phosphate buffered saline (PBS) and lysed in TNN lysis buffer (50mM Tris at ph 8.0, 120mM NaCl
More informationLightCycler Red 640-N-hydroxysuccinimide ester
For life science research only. Not for use in diagnostic procedures. R LightCycler Red 640-N-hydroxysuccinimide ester For labeling a minimum of 5 50 nmol oligonucleotides Content version: September 2016
More informationbeta-secretase Activity Assay Kit
beta-secretase Activity Assay Kit Catalog Number KA0900 100 assays Version: 04 Intended for research use only www.abnova.com Table of Contents Introduction... 3 Background... 3 General Information... 4
More informationDNA Hybridization and Detection
Chapter 6 DNA Hybridization and Detection Fluorescence Polarization Detection of DNA Hybridization........................................................ 6-2 Introduction.............................................................................................................
More informationCircLigase II ssdna Ligase
Cat. Nos. CL9021K and CL9025K www.lucigen.com MA298E-CircLigase II ssdna Ligase 2/2018 1 1. Introduction CircLigase II ssdna Ligase is a thermostable ligase that catalyzes iintramolecular ligation (i.e.,
More informationGenomic DNA Clean & Concentrator -25 Catalog Nos. D4064 & D4065
INSTRUCTION MANUAL Genomic DNA Clean & Concentrator -25 Catalog Nos. D4064 & D4065 Highlights Quick (5 minute) spin column recovery of large-sized DNA (e.g., genomic, mitochondrial, plasmid (BAC/PAC),
More informationProtocol for in vitro transcription
Protocol for in vitro transcription Assemble the reaction at room temperature in the following order: Component 10xTranscription Buffer rntp T7 RNA Polymerase Mix grna PCR DEPC H 2 O volume 2μl 2μl 2μl
More informationSupplementary Figures
Supplementary Figures Supplementary Figure 1. RCA reactions with C DNA i, r C DNA ii and rd2c1 using DP1 and DP2 as primers. (a) Sequence of rd2c1. It contains a linking duplex of 9 base pairs (boxed nucleotides);
More informationDNA 5 End-Labeling System
DNA 5 End-Labeling System I. Description...1 II. Product Components...2 III. Dephosphorylation Reaction...2 IV. Phosphorylation Reaction...3 V. Determination of Percent Incorporation/Specific Activity...3
More informationSupplementary information
Supplementary information ligonucleotide models of telomeric DA and RA form a hybrid G-quadruplex structure as a potential component of telomeres Yan Xu*, Takumi Ishizuka, Jie Yang, Kenichiro Ito, Hitoshi
More information5-methylcytosine enhance the substrate activity of DNA polymerase
5-methylcytosine enhance the substrate activity of DA polymerase Tian Tian, Shuang Peng, Heng Xiao, Yuelin Long, Boshi Fu, Xiaoe Zhang, Shan Guo, Shaoru Wang *, Xiang Zhou *, Songmei Liu, Xin Zhou Here,
More informationInstructions for Use Life Science Kits & Assays
Instructions for Use Life Science Kits & Assays Content Content 1 Product and order number... I 2 Storage conditions... I 3 Description... II 3.1 Quality data... II 3.2 Unit definition... II 4 Delivered
More informationFMF 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 information5.2 Protein purification
Purification of a His 6 -tagged Green Fluorescent Protein (GFP). Protein purification.. Purification of a His 6 -tagged Green Fluorescent Protein (GFP) Principle You can add either a N- or C-terminal His
More informationPROCEDURE FOR USE NICKEL NTA Magnetic Agarose Beads (5%)
1 AFFINITY HIS-TAG PURIFICATION PROCEDURE FOR USE NICKEL NTA Magnetic Agarose Beads (5%) DESCRIPTION Nickel NTA Magnetic Agarose Beads are products that allow rapid and easy small-scale purification of
More informationChIP-chip protocol adapted for the mod-encode project
ChIP-chip protocol adapted for the mod-encode project Version 1.2 : August 2007 Nicolas Nègre, Xiaochun Ni, Sergey Lavrov, Giacomo Cavalli and Kevin P. White University of Chicago, Department of Human
More informationMethods (detailed). for all the experiments. Cells were grown in 50 ml YEA. The cells were harvested and
Methods (detailed). Purification of yeast chromosomal DNA. Strain JZ105 (mat1m Δmat2,3::LEU2, ade6-210, leu1-32, ura4-d18, his2) was used for all the experiments. Cells were grown in 50 ml YEA. The cells
More informationTHUNDERBIRD SYBR qpcr Mix
Instruction manual THUNDERBIRD SYBR qpcr Mix 1304 A4251K THUNDERBIRD SYBR qpcr Mix QPS-201T 1 ml x 1 QPS-201 1.67 ml x 3 Contents [1] Introduction [2] Components [3] Primer design [4] Template DNA [5]
More informationProduct Specifications & Manual
Product Specifications & Manual Custom Oligo Synthesis, antisense oligos, RNA oligos, chimeric oligos, Fluorescent dye labeled oligos, Molecular Beacons, sirna, phosphonates Affinity Ligands, 2-5 linked
More informationWhy adapter ligation? Ligases. Oligonucleotide ligases. Definition of ligase
Why adapter ligation? Ligases Introduction to s in general, and RA 1 / RA 2, truncated in particular mira bacterial mra -P unknown sequence 3 -H -PPP unknown sequence 3 -H 3 adapter LIGASE catalyzed known
More informationElectronic Supporting Information
Electronic Supplementary Material (ESI) for ChemComm. This journal is The Royal Society of Chemistry 2015 Electronic Supporting Information Phosphorylation-Induced Hybridization Chain Reaction on Beads:
More informationSuperiorScript III cdna Synthesis Kit Instruction Manual
SuperiorScript III cdna Synthesis Kit Instruction Manual Cat.# EZ405S, EZ405M SuperiorScript III cdna Synthesis Kit Table of Contents I. Description... 3 II. Kit... 4 III. Procedure... 5 IV. Control Experiment
More informationStrep-Spin Protein Miniprep Kit Catalog No. P2004, P2005
INSTRUCTION MANUAL Strep-Spin Protein Miniprep Kit Catalog No. P2004, P2005 Highlights Fast protocol to purify Strep-tagged proteins from cell-free extracts Screen your recombinant colonies directly for
More informationMutagenesis PCR I (Multiple Site Directed Mutagenesis)
Mutagenesis Mutagenesis PCR I (Multiple Site Directed Mutagenesis) Mixture 25µl total reaction volume : 1. 2.5 µl of 10X Taq lligase buffer (need the NAD for Taq ligase) 2. 0.5 µl 100mM ATP 3. X µl (50-100
More informationSupporting Information for: DNA-based delivery vehicles: ph-controlled disassembly and cargo release by Jung-Won Keum and Harry Bermudez
Supporting Information for: DNA-based delivery vehicles: ph-controlled disassembly and cargo release by Jung-Won Keum and Harry Bermudez DNA sequences Strand Sequence 1- GGGTTAGGGTTAGGGTTAGGGAGGGTTAGGGTTAGGGTTAGGGAGGGTTAGGGTTAGGGTTAGGG
More informationStrep-Spin Protein Miniprep Kit Catalog No. P2004 & P2005
INSTRUCTION MANUAL Strep-Spin Protein Miniprep Kit Catalog No. P2004 & P2005 Highlights Fast & Simple: Purify Strep-tagged proteins from cell-free extracts using a spin-column in 7 minutes High-Quality:
More informationDNA Visualizer Extraction Kit
DNA Visualizer Extraction Kit Catalog Number D0006 50 reactions Version: 03 Intended for research use only www.abnova.com Table of Contents Introduction... 3 Intended Use... 3 Background... 3 General Information...
More informationDuplex-specific nuclease
Innovative Biotechnology Company www.evrogen.com Duplex-specific nuclease Product Cat.# Size Duplex-specific nuclease, lyophilized EA1 5 units* Duplex-specific nuclease, lyophilized EA2 units* Duplex-specific
More informationThe yield of transcripts for RNA polymerase regulated by hairpin structures in nascent RNA
Supporting Information The yield of transcripts for RNA polymerase regulated by hairpin structures in nascent RNA Satoru Nagatoishi a, Ryoya Ono b, Naoki Sugimoto a,b * a Frontier Institute for Biomolecular
More informationZR-96 Oligo Clean & Concentrator Catalog Nos. D4062 & D4063
INSTRUCTION MANUAL ZR-96 Oligo Clean & Concentrator Catalog Nos. D4062 & D4063 Highlights Quick, high-throughput (96-well) recovery of ultra-pure DNA and RNA oligonucleotides. Complete removal of dyes,
More informationGenomic DNA was extracted from 3 to 5 ml of blood collected in EDTA blood collection tubes
Supplementary information Methods DNA and RNA extraction Genomic DNA was extracted from to ml of blood collected in EDTA blood collection tubes using the Gentra Puregene Blood kit (Qiagen, California,
More informationSupporting Information. for. Angew. Chem. Int. Ed. Z Wiley-VCH 2003
Supporting Information for Angew. Chem. Int. Ed. Z52673 Wiley-VCH 2003 69451 Weinheim, Germany Modular Assembly of Glycoproteins: Towards the Synthesis of GlyCAM-1 Using Expressed Protein Ligation. D.
More informationSupporting Information
Supporting Information One-step, Multiplexed Fluorescence Detection of micrornas Based on Duplex-Specific Nuclease Signal Amplification Bin-Cheng Yin, Yu-Qiang Liu, and Bang-Ce Ye* Lab of Biosystems and
More informationAmplification Products for PCR and RT-PCR
Selection guide Polymerase Hot start Comment UptiTherm DNA pol. no Most economic. Lower error rate than Taq polymerase Available in several formats, master mix including or not dntp, Mg 2+..., in gel format
More informationMirror-image polymerase chain reaction
Supplementary Information Mirror-image polymerase chain reaction Wenjun Jiang 1,4, Baochang Zhang 2,4, Chuyao Fan 1,4, Min Wang 1,4, Jiaxing Wang 2, Qiang Deng 1, Xianyu Liu 1, Ji Chen 1, Jishen Zheng
More informationElectronic Supporting Information
Electronic Supplementary Material (ESI) for Chemical Science. This journal is The Royal Society of Chemistry 2017 Electronic Supporting Information Highly Sensitive and Multiplexed Quantification of mrna
More informationGenomic DNA Clean & Concentrator -10 Catalog Nos. D4010 & D4011
INSTRUCTION MANUAL Genomic DNA Clean & Concentrator -10 Catalog Nos. D4010 & D4011 Highlights Quick (5 minute) spin column recovery of large-sized DNA (e.g., genomic, mitochondrial, plasmid (BAC/PAC),
More information3color RT HS-PCR Mix SYBR Ready-to-use mix for real-time Hot Start PCR with SYBR Green. Dedicated for white reaction tubes and plates.
3color RT HS-PCR Mix SYBR Ready-to-use mix for real-time Hot Start PCR with SYBR Green. Dedicated for white reaction tubes and plates. version 0217 250 reactions in 20 μl Cat. # 2000-250S 2500 reactions
More information7 Synthesizing the ykkcd Mutant Toxin Sensor RNA in vitro
7 Synthesizing the ykkcd Mutant Toxin Sensor RNA in vitro 7.1 Learning Objective In the quest toward understanding how the ykkcd toxin sensor recognizes the antibiotic tetracycline you thus far designed
More informationProteasome Activity Fluorometric Assay Kit II (Cat. # J4120)
Proteasome Activity Fluorometric Assay Kit II (Cat. # J4120) Each supplied substrate is sufficient for use in 250 X 100 µl reactions to monitor the chymotrypsin-like (Suc-LLVY- AMC), trypsin-like (Boc-LRR-AMC)
More informationSolid Phase cdna Synthesis Kit
#6123 v.02.09 Table of Contents I. Description... 2 II. Kit components... 2 III. Storage... 2 IV. Principle... 3 V. Protocol V-1. Preparation of immobilized mrna... 4 Protocol A: Starting from Tissue or
More informationCloning 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 informationAmpliScribe T7-Flash Biotin-RNA Transcription Kit
AmpliScribe T7-Flash Biotin-RNA Transcription Kit Cat. No. ASB71110 Available exclusively thru Lucigen. lucigen.com/epibio www.lucigen.com MA276E AmpliScribe T7-Flash Biotin-RNA Transcription Kit 12/2016
More informationPrepare a Barcoded Fragment Library with the SOLiD Fragment Library Barcoding Kit 1 96
QUICK REFERENCE CARD Prepare a Barcoded Fragment Library with the SOLiD Fragment Library Barcoding Kit 1 96 Note: For safety and biohazard guidelines, refer to the Safety section in the Applied Biosystems
More informationPurification and Sequencing of DNA Guides from Prokaryotic Argonaute Daan C. Swarts *, Edze R. Westra, Stan J. J. Brouns and John van der Oost
Purification and Sequencing of DNA Guides from Prokaryotic Argonaute Daan C. Swarts *, Edze R. Westra, Stan J. J. Brouns and John van der Oost Department of Agrotechnology and Food Sciences, Wageningen
More informationRNA-direct Realtime PCR Master Mix
Instruction manual RNA-direct Realtime PCR Master Mix 0803 F0929K RNA-direct Realtime PCR Master Mix Contents [1] Introduction [2] Components [3] Primer/Probe design [4] Detection [5] Specimens [6] Protocol
More informationab Oligonucleotide Conjugation Kit
ab188289 Oligonucleotide Conjugation Kit Instructions for Use For the Covalent Conjugation of Antibodies and Oligonucelotides. This product is for research use only and is not intended for diagnostic use.
More informationQS S Assist STK_FP Kit
QS S Assist STK_FP Kit Description STK FP kit is designed for use in pharmacological assays for STK based on fluorescence polarization. The kit includes assay buffer, human protein kinase, ATP/fluorescence-
More informationSupporting Information
Supporting Information Chemical Modification-Assisted Bisulfite Sequencing (CAB-Seq) for 5-Carboxylcytosine Detection in DNA Xingyu Lu 1, Chun-Xiao Song 1, Keith Szulwach 2, Zhipeng Wang 1, Payton Weidenbacher
More informationG-Quadruplex formation using fluorescent oligonucleotide as a detection method for discriminating AGG trinucleotide repeats
Electronic Supplementary Material (ESI) for ChemComm. This journal is The Royal Society of Chemistry 1 Electronic Supplementary Information G-Quadruplex formation using fluorescent oligonucleotide as a
More informationRP 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 informationNanoparticle-Based Bio-Bar Codes for the Ultrasensitive Detection of Proteins. Jwa-Min Nam, C. Shad Thaxton, and Chad A. Mirkin
1 Nanoparticle-Based Bio-Bar Codes for the Ultrasensitive Detection of Proteins Jwa-Min Nam, C. Shad Thaxton, and Chad A. Mirkin Department of Chemistry and Institute for Nanotechnology, Northwestern University,
More informationGeneration of gene knockout vectors for Dictyostelium discoideum
Generation of gene knockout vectors for Dictyostelium discoideum Instruction manual Last date of revision April 2012 2015 Version PR29-0001 PR29-0003 www.stargate.com This manual can be downloaded under
More informationTable of contents. I. Description...2. II. Principle...2. III. Kit Components...3. IV. Storage...3. V. Features...4. VI. Precautions for Operation...
Table of contents I. Description...2 II. Principle...2 III. Kit Components...3 IV. Storage...3 V. Features...4 VI. Precautions for Operation...4 VII. Protocol...4 VIII.Experiment Example...6 IX. Appendix...8
More informationSensoLyte AMC Calpain Activity Assay Kit *Fluorimetric*
SensoLyte AMC Calpain Activity Assay Kit *Fluorimetric* Catalog # 72150 Kit Size 100 Assays (96-well plate) Optimized Performance: This kit is optimized to detect calpain activity Enhanced Value: It provides
More informationHE Swift Cloning Kit
HE Swift Cloning Kit For high-efficient cloning of PCR products either blunt or sticky-end Kit Contents Contents VTT-BB05 phe Vector (35 ng/µl) 20 µl T4 DNA Ligase (3 U/µl) 20 µl 2 Reaction Buffer 100
More informationGuide-it sgrna In Vitro Transcription and Screening Systems User Manual
Guide-it sgrna In Vitro Transcription and Screening Systems User Manual Cat. Nos. 632638, 632639, 632635, 632636, 632637 (040618) 1290 Terra Bella Avenue, Mountain View, CA 94043, USA U.S. Technical Support:
More informationMMLV 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 informationSupporting Information. Quencher Group Induced High Specificity Detection of. Telomerase in Clear and Bloody Urines by AIEgens
Supporting Information Quencher Group Induced High Specificity Detection of Telomerase in Clear and Bloody Urines by AIEgens Yuan Zhuang, a Mengshi Zhang, a Bin Chen, c Ruixue Duan, a Xuehong Min, a Zhenyu
More informationRealHelix TM qrt-pcr Kit [Intercalator type]
RealHelix TM qrt-pcr Kit [Intercalator type] CERTIFICATE OF ANALYSIS (1603-V01R03) Kit contents RealHelix TM qrt-pcr Kit [Intercalator type] Cat. No. QRT-S100 (100 rxns) QRT-S500 (500 rxns) qrt-pcr Enzyme
More informationCell-Free Protein Expression Kit
Cell-Free Protein Expression Kit Handbook Version v.01, January 2018 Cat# 507024 (Sigma 70 Master Mix Kit, 24 Rxns) Cat# 507096 (Sigma 70 Master Mix Kit, 96 Rxns) Please refer to this product in your publication
More informationFor Research Use Only Ver
INSTRUCTION MANUAL Oligo Clean & Concentrator Catalog Nos. D4060 & D4061 Highlights Quick (2 minute) recovery of ultra-pure DNA and RNA oligonucleotides. Complete removal of dyes, salts, enzymes, nucleotides,
More informationSingle tube gene synthesis by phosphoramidate chemical ligation Supplementary Information
Electronic Supplementary Material (ESI) for Chemical Communications. This journal is The Royal Society of Chemistry 2017 Single tube gene synthesis by phosphoramidate chemical ligation Supplementary Information
More informationGuide-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 informationSYBR 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 informationSupporting Information. Synthesis and evaluations of an acid-cleavable, fluorescently labeled
Electronic Supplementary Material (ESI) for ChemComm. This journal is The Royal Society of Chemistry 2015 Supporting Information Synthesis and evaluations of an acid-cleavable, fluorescently labeled nucleotide
More informationSOP for Quantitation of the Hepatitis C Virus (HCV) Genome by RT-PCR
Virus Bank SOP-HCV-001 1. Scope SOP for Quantitation of the Hepatitis C Virus (HCV) Genome by RT-PCR 1.1 This procedure describes a method for quantitation of the HCV genome in HCV-infected cells by RT-PCR.
More informationElectronic Supplementary Information (ESI)
Electronic Supplementary Material (ESI) for RSC Advances. This journal is The Royal Society of Chemistry 2015 Electronic Supplementary Information (ESI) A label free fluorescent assay for uracil-dna glycosylase
More informationSupporting Information
Supporting Information Wiley-VCH 2005 69451 Weinheim, Germany Calderone and Liu Supporting Information page 1 Small-Molecule Diversification From Iterated Branching Reaction Pathways Enabled by DNA-Templated
More information