Agarosegelelectrophoresis
General ProcedureCast a gelPlace it in gel box in running bufferLoad samplesRun the gelImage the gelCasting Gels0.7% agarose gel with 1kbp ladder in UV and white light showing different dyesThe amount of agarose to use in your gel depends on the DNA in question. Use the following table as a rough guide:Agarose Concentration (g/100mL)Optimal DNA Resolution (kb)0.51 - 300.70.8 - 121.00.5 - 101.20.4 - 71.50.2 - 3Measure......阅读全文
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
Thermal－Inactivation
Thermal InactivationA simple, reversible way to a stop restriction reaction is by adding EDTA, which chelates Mg2+, thereby preventing catalysis. If f
General－Cloning－Protocols
Large Scale Preps: (See Large scale plsasmid prep protocol for more details)Cultures: Inoculate a 5 mL LB/Amp (50 - 100 µg/mL) culture in early a.m. w
Apoptosis:－A－Laboratory－Manual－of－Experimental－Methods－Andrea－Cossarizza
THE CELL 1. Morphological aspects of apoptosis Walter Malorni, Stefano Fais & Carla Fiorentini 2. Cell cycle Miriam Capri & Daniela BarbieriTHE NUCLEU
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
Construction－of－BAC－Libraries：Megabase－DNA－Isolation
Megabase DNA IsolationMegabase-size DNA isolation from plantsTo construct large insert DNA libraries in BAC and YAC vectors, methods must be developed
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
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
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
An－Integrative－Procedure－for－Apoptosis－Identification－and－Measurement2
TroubleshootingCritical Steps(1) Don’t trypsinize cells for too long when collecting them.(2) Rotation speed should be no more than 1500 rpm during ce
DNA转化实验指导5
2C. Notes on Transformation 1. Bacto tryptone and yeast extract can cause allergic reactions. 2. All containers used to handle the bacteria (
蛋白质分析技术(Analytical－Techniques－for－Protein)2
二、透析和超滤1.透析（Dialysis）：就是利用蛋白质分子不能通过半透膜（Semipermeable membrane）而小分子可以自由透过的性质，使蛋白质与小分子物质分开。作用：脱盐、无机离子和小分子物质等。透析膜：动物膜、羊皮纸、火棉胶、赛璐玢或其他材料等。2.超滤（ultrafiltrat
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
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
重组DNA的分离、克隆与测序实验手册4
H. Fragment purification on Sephacryl S-500 spin columnsDNA fragments larger than a few hundred base pairs can be separated from smaller fragments by
A－simple,－rapid－procedure－for－the－isolation－of－DNA－for－PCR
The polymerase chain reaction (PCR) is a method for amplifying specific segments of DNA defined by the small primers used to start the reaction. Using
Quantitative－PCR
实验概要Quantitative PCR involves co-amplification of two templates: a constant amount of a preparation containing the desired target sequence and var
DNA抽提
DNA抽提（主要内容如下）· Working with DNA· DNA Extraction from Bacteria and Other Organisms· DNA Extraction from Cell and Tissue· Mitochondria DNA Isola
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
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
CORE－SAMPLE－PCR:－A－method－to－rePCR－unique－bands－from－products－of－mixed－s
INTRODUCTIONThe products of a PCR reaction - especially when this is done on eukaryotic genomic DNA, and when using degenerate primers - often contain
定量PCR实验技术－QPCR
Quantitative PCRJoseph SambrookPeter Maccallum Cancer Institute and The University of Melbourne, AustraliaDavid W. RussellUniversity of Texas Southwes
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
CORE－SAMPLE－PCR
A method to re-PCR unique bands from products of mixed sizeContentsINTRODUCTIONPROTOCOLCOMMENTSINTRODUCTIONThe products of a PCR reaction - especially
A－simple,－rapid－procedure－for－the－isolation－of－DNA－for－PCR
N.M. DuTeau and J.F. Leslie - Department of Plant Pathology, Throckmorton Hall, Kansas State University, Manhattan, KS 66506-5502The polymerase chain
An－Integrative－Procedure－for－Apoptosis－Identification－and－Measurement
IntroductionApoptosis is a normal physiological phenomenon put forward by Kerr [1]. It plays an important role in embryonic development, maintenance o
Ligation－Optimization
The following protocol can be used to optimize ligation conditions for difficult to clone (e.g. very large) fragments. The principle is to independent