SingletubeconfirmationPCRprotocol
The following colony PCR protocol has been designed to be performed in individual reaction tubes. We usually test three colonies from each transformation along with a wild-type control. A series of five different PCR tests are performed on each colony to confirm both of the novel recombination junctions .1. Clonal purification- Pick three colonies from a transformation plate and streak each one out to single colonies......阅读全文
Single－tube－confirmation－PCR－protocol
The following colony PCR protocol has been designed to be performed in individual reaction tubes. We usually test three colonies from each transformat
PCR－protocol
PCR reactionProtocol for 50µl reaction - adjust amounts if necessary, for a 20µl reaction use the same volumes of primer and dNTP-mix, but adjust the
酵母遗传学技术
Genome-wide Gene Expression Analysis (Richard Young Research Group,Whitehead Institute for Biomedical Research)A genoe-wide gene expression analysis u
Detection－of－Viruses－in－Infected－Plant－Extracts－using－ImmunocapturePCR
1) Immunocapture stageCoating buffer: 15 mM Na2CO3; 35 mM NaHCO3, and 3 mM NaN3, per liter (pH 9.6).Extraction buffer: (20 mM Tris-HCL (pH 8.0), 138
Detection－of－Viruses－in－Infected－Plant－Extracts－using－ImmunocapturePCR
1) Immunocapture stageCoating buffer: 15 mM Na2CO3; 35 mM NaHCO3, and 3 mM NaN3, per liter (pH 9.6).Extraction buffer: (20 mM Tris-HCL (pH 8.0), 138
其它PCR方法
· Standard PCR Protocol (Molecular Biology Techniques Manual)The followings are described in detailRecommended Reagent ConcentrationsRecommend
Colony－PCR－Protocol
1. Pull out eight glycerol stock plates from the –80oC freezer and set on bench top to thaw. Be sure to remove the foil seal before leaving the plates
TAIL－PCR－Protocol
TAIL is a series of reactions that are intended to map where a T-DNA (transfer DNA) has inserted within the genome. The main components of the 3 react
Single－Cell－PCR－单细胞PCR实验技术
Single Cell PCR(Protocol provided by Carolyn Troeger)Cell picking c Axiovert 100/Zeiss, extended glass capillary/Drummond, Broomall and a micromanipul
Single－Primer－(SemiRandom)－PCR
DescriptionSingle primer PCR allows amplification from known to unknown regions in chromosomes, phage, plasmids, large PCR products and other sources
RTPCR－PROTOCOL
RT-PCR PROTOCOL材料与方法………………………………………………………… 1．材料 ………………………………………………………1.1 供试用组织(细胞)…………………………………1.2 主要仪器设备………………………………………1.3 主要试剂……………………………………………1.
SYBR－Green－Quantitative－PCR－Protocol
SummaryQuantitative PCR is a method used to detect relative or absolute gene expression level. All qPCR involves the use of fluorescence to detect the
Protocol－for－competitive－RTPCR
For quantifying mRNA, we use a competitive RT-PCR protocol with internal standard RNAs. These are added in a defined quantity to the RNA sample prior
The－protocol－for－LIC－by－Exonuclease－III
The protocol for LIC by Exonuclease III梁耀极1. Design the primers with 15-bp overlap;2. Digest the vector by proper restriction enzyme;For getting high
基于PCR技术的染色质沉淀分析2
METHOD Analysis of precipitated chromatin fractions (from Chromatin Immunoprecipitation on Unfixed Chromatin from Cells and Tissues to Analyze Histone
Tube－formation－assay
DescriptionThis is a fast and easy assay to test the angiogenic/anti-angiogenic properties of molecules. As compared to other angiogenesis assays, suc
Sitedirected－Mutagenesis－using－PCR
Site-directed Mutagenesis using PCRMichael P. Weiner, Tim Gackstetter, Gina L. Costa, John C. Bauer, and Keith A. KretzFrom: Molecular Biology: Curren
Basic－PCR
实验概要The following basic protocol serves as a general guideline and a starting point for any PCR amplification. Optimal reaction conditions (incubati
Protocol－for－dsRNA－Synthesis
实验概要 We routinely produce dsRNA by in vitro transcription of a PCR generated DNA template containing the T7 promoter sequence on both ends
Vacuum/Spin－Protocol－for－Tissue－DNA－Extraction
实验概要The E.Z.N.A.® Tissue DNA Kit provides a rapid and easy method for the isolation of genomic DNA for consistent PCR and Southern analysis. Up to 3
Bacterial－Colony－PCR
Bacterial Colony PCRObjective:This protocol allows rapid detection of transformation success when primers are available to allow determination of corr
qPCR－Protocol－for－SNP－Genotyping
实验概要Platinum® qPCR SuperMix for SNP Genotyping is a ready-to-use reaction mix for the amplification and identification of single-nucleotide polymorp
Protocols－for－LCM－preparation－and－analysis
Protocols for LCM preparation and analysis I. Preparation, LCM and RNA/DNA extraction of Frozen Tissue SectionsA. EmbeddingB. CuttingC. StainingII. Pr
E.Z.N.A.®－Plasmid－Maxi－Kit－Spin－Protocol
实验概要This Protocol is designed to isolate 500-1200 ug of high Copy-Number plasmids or 50-400 ug of low Copy-Number Plasmids from 200 ml overnight c
REALTIME－PCR－WITH－SYBR－GREEN－I－PROTOCOL
1、反应体系 25ulCocktail 20 ul: Primer FP 1 ul + Primer RP 1u + 2 * SYBR GREEN I MIX 12.5 + H2O 5.5 ul cDNA 5 ul2、Primer 浓度，需要摸索，从200nM到700nm等，设置不同浓度。3、每个引
E.Z.N.A.®－Protocol－for－ParaffinEmbedded－Tissue
实验概要The E.Z.N.A.® Tissue DNA Kit provides a rapid and easy method for the isolation of genomic DNA for consistent PCR and Southern analysis. Up to
E.Z.N.A.®－Plasmid－Maxi－Kit－vacuum－Protocol
实验概要This Protocol is designed to isolate 500-1200 ug of high Copy-Number plasmids or 50-400 ug of low Copy-Number Plasmids from 200 ml overnight c
How－do－you－synthesize－your－dsRNA
We routinely produce dsRNA by in vitro transcription of a PCR generated DNA template containing the T7 promoter sequence on both ends (I. Primer Desig
果蝇RNAi的实验中双链短RNA的合成（dsRNA)方法
实验概要We routinely produce dsRNA by in vitro transcription of a PCR generated DNA template containing the T7 promoter sequence on both ends (I. Prim
Screening－BAC－filters－with－nonradioactive－probes
(Protocol for a single high-density BAC colony filter (HDR filter) of 22 cmx22 cm) to be screened using Amersham''s ECL hybridization kit; thi