Standardneutralagaroseelectrophoresis
Standard neutral agarose electrophoresisStandard agarose gels can be prepared using either TBE or TAE running buffers.You will need:Either 10 x TBE or 10 x TAE buffers1 x TBE buffer - 89mM Tris, 89mM boric acid, 2mM EDTA, pH 8.1 x TAE buffer - 40mM Tris-acetate, 2mM EDTA, pH 8.10mg/ml ethidium bromide stock solution10 x Type III loading buffer (50% glycerol, 0.5% bromophenol blue, 0.5% xylene cyanol, this solution sh......阅读全文
Basic－procedures－for－bacteria－culture1
A. Phenol extraction of DNA samplesPhenol extraction is a common technique used to purify a DNA sample (1). Typically, an equal volume of TE-saturated
Standard－PCR－reaction
Steps for Standard PCR ReactionDesign primers. In general, primers should have the following properties:Tip: Primer3 is an excellent resource for choo
Construction－of－BAC－Libraries：Construction－of－a－BAC－library
Once high molecular weight (HMW) DNA has been prepared it must somehow be fragmented and DNA in the desired size range isolated. In general, as the de
DNA－labeling－by－nick－translation
DNA labeling by nick translationreagents: DNA for labeling (concentration c > 150 ng/µl) modified nucleotides: Biotin-16-dUTP, Digoxigenin-11-dUTP, co
Standard－RTPCR
RT-PCR or reverse transcription PCR refers to PCR that uses product of an RT reaction as template. In effect, the PCR amplifies cDNA fragments. In one
Thermal－Inactivation
Thermal InactivationA simple, reversible way to a stop restriction reaction is by adding EDTA, which chelates Mg2+, thereby preventing catalysis. If f
Purifying－Large－E.－coli－Restriction－Fragments－from－PulsedField－Gels
DNA PreparationE. coli chromosomal DNA is prepared following the method of Heath et al. ( J. Bacteriol., 174, 1992). Cells are embedded in agarose, th
electrophoresis－of－DNA
Agarose Gel Electroporesis of DNA Making the gel: 1. Place casting platform with well former sideways in gel stand where you wish to pour
Capillary－Electrophoresis
Capillary electrophoresis is a very sensitive analytical technique. Sample components are separated within a fused silica capillary using one of sever
Chromatin－Electrophoresis
Dr. William H. Heidcamp, Biology Department, Gustavus Adolphus CollegeExercise 10.4 - Chromatin ElectrophoresisLEVEL IIMaterials 14 M Urea6 M NaCl0.05
Protein－Electrophoresis
DefinitionAmino acids, nucleotides, polypeptides, and other compounds in a colloidal state can be separated by the application of external voltages wh
DNA的酶学操作
DNA的酶学操作DNA Modifying Enzymes (Michael Blaber)Introduction to bacterial restriction/modification system. It provides very useful background knowledge
Facts－and－trouble－shooting
10 Fun Facts for DNA Electrophoresis::Migration of DNA is retarded and band distortion can occur when too much buffer covers the gel. The slower migra
Top－10－Fun－Facts－for－DNA－Electrophoresis
Did you know:When preparing agarose for electrophoresis, it is best to sprinkle the agarose into room-temperature buffer, swirl, and let sit at least
DNA电泳（agarose胶）
DNA琼脂糖凝胶电泳 实验方法原理 利用DNA分子在琼脂糖凝胶中泳动时具有的电荷效应和分子筛效应。电荷效应是指DNA分子在高于等电点的pH溶液中带负电，在电场
DNA电泳（agarose胶）
实验材料 DNA样品试剂、试剂盒 琼脂糖 电泳缓冲液溴化乙锭 上样缓冲液仪器、耗材 电泳仪电泳漕 透射紫外灯 胶带纸 紫外成像仪
DNA电泳（agarose胶）
DNA电泳可用于：（1）分离不同大小的DNA片段；（2）鉴定目的DNA片段；（3）纯化和回收DNA片段。实验方法原理利用DNA分子在琼脂糖凝胶中泳动时具有的电荷效应和分子筛效应。电荷效应是指DNA分子在高于等电点的pH溶液中带负电，在电场中向正极移动，且相同数量的双链DNA几乎具有等量的净电荷，能以
重组DNA的分离、克隆与测序实验手册8
B. Midiprep double-stranded DNA isolationA midi-prep double-stranded DNA isolation has been developed to generate a sufficient amount of template DNA
Gel－Electrophoresis－of－DNA
What is Electrophoresis?Electrophoresis is a technique used in the laboratory that results in the separation of charged molecules. In this CyberLab we
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
RNA－gel－electrophoresis
实验概要RNA gel electrophoresis主要试剂DEPC H2ODEPC 0.1% (v/v)q.s. de-ioinized H2O37ºC x1 hr, or r.t. overnightAutoclave.(NaOAc, EDTA and ethidium bromide sol
RNA－gel－electrophoresis
MaterialsDEPC H2ODEPC 0.1% (v/v)q.s. de-ioinized H2O37ºC x1 hr, or r.t. overnightAutoclave.(NaOAc, EDTA and ethidium bromide solutions should also be
基于PCR技术的染色质沉淀分析1
INTRODUCTIONAfter chromatin immunoprecipitation (ChIP), different PCR-based approaches can be used to determine how much DNA is precipitated at a locu
Preparation－of－Agarose－Gels－for－DNA－separations
Weigh out the desired amount of agarose and place in an Erlenmeyer flask with a measured amount of electrophoresis buffer, e.g. for an 0.8% gel, add 0
Agarose－Gels－for－Single－Stranded－DNA
1. Prepare 50X TAE as:242 g Tris Base57.1 mL Glacial Acetic Acid100 mL 500 mM EDTA, pH 8.0600 mL ddH2OMix. Bring volume to 1 L. Autoclave.2. Mix the f
The－UnderAgarose－Migration－Assay
overviewThe Under-Agarose assay is a useful method for observing the response of a cell population to one or more chemoattractant sources. The behavio
DNA－Purification－from－Agarose－Gels
1. Separate DNA fragments in an agarose gel cast with 0.5 mg/mL Ethidium bromide. Locate bands with a hand-held long-wave UV lamp.2. Slice the gel wit
Standard－Protocols－Autoradiography－(35S)
Remove Kodak NTB2 nuclear emulsion from fridge and place at 42oC for around 30-60 mins (until melted).Make up the developer and the fixer and place in
基于PCR技术的染色质沉淀分析
INTRODUCTION After chromatin immunoprecipitation (ChIP), different PCR-based approaches can be used to determine how much DNA is precipitated at a loc
质粒的小量制备
· Standard (alkaline lysis) Mini-Prep (Goldberg Lab)Standard protocol for mini-prep and recipe for solution I, II and III.Alkaline Lysis Minip