RestrictionDigestsofHighMolecularWeightYeastDNA
Purpose:To perform restriction digests of YACs for mapping using rare cutting enzymes or more conventional restriction endonuclease digestion.Time required:2-4 hoursSpecial Reagents:dependent upon enzymes desiredProcedure:Using wide-bore pipette, aliquot 100 ng of liquid DNA into eppendorf tube. Add 2.5 祃 10X restriction buffer, 1-2 ul RNase (5 mg/ml stock), 250-500 ng BSA, then dH2O up to 25 祃 volume. Incubate at ap......阅读全文
2d2D电泳
For an in-depth review of the method, see Friedman, K. and B. Brewer (1995) Analysis of replication intermediates by two-dimensional agarose g
Preparation－of－Sonicated－Human－DNA
Purpose:To break up high molecular weight human placental DNA into fragment sizes of 500 bp or less which can be used as competitor DNA in Southern an
用超声波处理人类基因组DNA
Purpose:To break up high molecular weight human placental DNA into fragment sizes of 500 bp or less which can be used as competitor DNA in Southern an
DNA转化实验指导2
1B. Cloning 1. A caveat on dephosphorylation: the most common reason for failure to obtain colonies is a result of adding too much BAP or CIP to
Thermal－Cycling－Profile－for－Standard－PCR
Initial denaturationIt is very important to denature the template DNA completely. Initial heating of the PCR mixture for 2 minutes at 94°–95°C is enou
线粒体荧光探针大全：TMRM，Mitotracker，JC1（5）
Monoclonal Antibodies Specific for Proteins in the Oxidative Phosphorylation SystemOxidative phosphorylation (OxPhos) activity occurs in the mitochond
三代测序的DNA提取和宏基因组学分析（一）
改进的人类肠道微生物组的高分子量DNA提取，纳米孔测序和宏基因组学装配Improved high-molecular-weight DNA extraction,nanopore sequencing and metagenomicassembly from the human gut microb
The－ribonuclease－protection－assay－(RPA)
The ribonuclease protection assay (RPA) is a highly sensitive and specific method for the detection of mRNA species. The assay was made possible by th
细菌的核酸抽提
DNA Extraction· DNA Extraction from Bacteria (Julie B. Wolf,UMBC)Phenol/chloroform method· DNA Extraction From Bacteria (Triton Method
DNA－Isolation－From－BAC－－PAC－Clones
DNA Isolation From BAC & PAC Clones This is a rapid alkaline lysis miniprep method for isolating DNA from large PAC clones. It is a modification of a
DNA标记
DNA标记（主要内容如下） DNA Labeling by Nick Translation Random Primed Labeling End-Labeling Purification of Labeled DNA Non-isotopic Labeling OthersDNA L
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
RNA实验方法3
Part III: HybridizationTurn heatblock on to 95C.For samples in water or ethanol, dry down appropriate amount of RNA, and include a tube with 1ul of tR
Transformation－of－Magnaporthe－grisea－to－phosphinothricin－resistance
Three transformation systems have been reported for the rice blast fungus Magnaporthe grisea (Parsons et al. 1987 Proc. Natl. Acad. Sci. USA 84:4161-4
其它PCR方法
· Standard PCR Protocol (Molecular Biology Techniques Manual)The followings are described in detailRecommended Reagent ConcentrationsRecommend
Standard－neutral－agarose－electrophoresis
Standard neutral agarose electrophoresisStandard agarose gels can be prepared using either TBE or TAE running buffers.You will need:Either 10 x TBE or
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
SOUTHERN－BLOTTING
Materials:Whatman 3 mm Blotting Papernitrocellulose (Schleicher & Schuell, Amersham) or nylon membrane filter (Amersham).Paper towels (preferably C-fo
DNA转化实验指导1
CONTENT Transformation-Competent E. coli preparation Inoue "ultra-competent" methodRubidium chloride methodCosmid packaging protocol DNA Ligation an
Yeast－Cell－Cycle－by－Flow－Cytometry
ReagentsCold absolute ethanol.0.5 M Na citrate stock (filtered), 50mM diluted stock.10 mg/ml RNase A (Boil 10 mins, cool, filter and store at -20°C).4
Acid－Phenol－Yeast－RNA－Prep
This is the preferred method for yeast RNA preparationuse Gloves and RNAse free solutions throughout.1. Use a YPD overnight culture to innoculate fres
High－Resolution－Agarose－Gel－Electrophoresis
实验概要Agarose gel electrophoresis remains the most widely used technique for separating nucleic acid fragments due to its ease of use, non-toxicity, a
Fungal－Genomic－DNA－Extraction
OverviewHigh throughput of many fungal isolates can be achieved by growing axenic cultures in either (a) 1.5mL microfuge tubes, half full with liquid
Fungal－Genomic－DNA－Extraction
OverviewHigh throughput of many fungal isolates can be achieved by growing axenic cultures in either (a) 1.5mL microfuge tubes, half full with liquid
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
Mitochondrial－DNA－Isolation－from－Somatic－Embryogenic－Cell－Cultures－of－Larix
Mitochondrial DNA is isolated by a modification of the methods described by Wilson and Chourey (1984) and Radetzky (1990). Cell cultures at four days
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
DNA－Immunoprecipitation－for－the－Determination－of－DNABinding－Specificity
Andrea J. Gossett and Jason D. Lieb1Department of Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA1Corresponding autho
UV－CrossLinking－an...－（三）
9. Gel purification of cDNA 1) Spin down and wash the samples (see 8.1), then resuspend the pellets in water (6 μl) 2) Add 2x TBE-urea loading b
Cellular－－Molecular－Pathology－Branch
VisionTo provide scientific collaboration of excellence to National Toxicology Program (NTP) (http://ntp.niehs.nih.gov/) interdisciplinary research pr