HighResolutionAgaroseGelElectrophoresis
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, and broad separation range. By varying agarose concentration, gel pore size can be controlled to separate nucleic acid molecules in a wide range of sizes. The migration of nucleic acids in agarose gels is affected by the choice of buffer......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
AMRESCO琼脂糖的使用方法指导
正确的加热方式对琼脂糖的充分溶解和凝结至关重要，在此，我们提供以下几点作为正确制备琼脂糖凝胶的操作参考。*AMRESCO琼脂糖加热溶解所需时间短。*琼脂糖在TAE或者TBE缓冲液中不溶解，加热时，琼脂糖颗粒水化形成溶液，水化是时间依赖的，不同的琼脂糖水化点不同。AMRESCO琼脂糖纯度高，超微颗粒，
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
Northern－Blotnorthern杂交
Northern BlotPreparation of Formaldehyde Agarose GelThe gel conditions (1% agarose, 1X MOPS, 6.3% formaldehyde) are designed for ~4 hours of electroph
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
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
Determining－the－Direction－of－Replication－Fork－Movement
For an in-depth review of the method, see Friedman, K. and B. Brewer (1995) Analysis of replication intermediates by two-dimensional agarose gel elect
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
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
DNA－mobility－in－gels
1. Migration of marker dyes in native polyacrylamide non-denaturing gels Gel % Bromophenol blue (BP) Xylene cyanole (XC) 3.5 100 460 5.0
Pulse－Field－Electrophoresis
Manipulating and analyzing DNA are fundamentals in the field of molecular biology. Indeed, separating complex mixtures of DNA into different sized fra
Construction－and－Manipulation－of－LargeInsert－Bacterial－Clone－Libraries
Acknowledgements The organizer of the workshop acknowledges Dr. Murray Milford, Professor and Interim Head, and Dr. Mark Hussey, Professor and Interim
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
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
Methylene－Blue－DNA－staining－protocol
Methylene Blue DNA staining protocolProtocol:Load 2-5X the amount of DNA that would give bands of moderate intensity on an ethidium bromide stained ge
Silver:－TimeLapse－Microscopy
Pad Preparation1. Microwave 2% agarose (mix of low-melt and normal, to taste) in Thorn media (see below). (If you have used different percentages of a
Transplantation－of－theeyeforming－region－of－Amphibian－neurula－into－theflank
EMBRYO CELL AND TISSUE CULTURE TECHNIQUESRearing solution: 10% Modified Steinberg's Solution. Operating solution:Full strength HBSt solution.Micro
实验方法丨蛋白质体外瓜氨酸化
组蛋白的翻译后修饰过程对 DNA 转录，染色体功能以及维护细胞信号传导通路的正常运行具有重要的作用，瓜氨酸化是由称为蛋白质精氨酸脱亚胺酶（Protein Arginine Deiminases，PADs）的酶家族催化的许多关键组蛋白修饰手段之一。PADs 催化肽基精氨酸水解，在组蛋白上形
CORE－SAMPLE－PCR
A method to re-PCR unique bands from products of mixed sizeContentsINTRODUCTIONPROTOCOLCOMMENTSINTRODUCTIONThe products of a PCR reaction - especially
蛋白质电泳
蛋白质电泳（主要内容如下）One-Dimensional SDS-PAGETwo-Demensional SDS-PAGEProtein Electrophoresis in Agarose Gel Gel StainingRecipesOne-Dimensional SDS-PAGE·
乙酰化蛋白快速检测和定量
Rapid detection and quantitation of total cellular acetylated proteins using our research products
In－vitro－growth－of－seedlings
sterilisation of seeds: rinse with 70% EtOH for 30 sec put in 1% bleach (sodium hypochlorite, supplemented by a few drops of Tween-20) for 5 mi
DNA－laddering－assay－for－treated－cells
Characteristics of this procedure:I found the procedure described by Gong et al. to be a convenient and successful method to detect DNA laddering in c
免疫共沉淀
实验概要本实验以植物叶片为试材介绍了免疫共沉淀的基本方法。主要试剂50uM雌激素，液氮，提取缓冲液TBS(50mM Tris-Cl，0.15M NaCl，pH 7.4，1mM PMSF，5mM Benzamidine)，glycine-HCl，TBS，洗脱缓冲液(TBS，0.5mg/mL 3
免疫共沉淀概述
实验概要本实验以植物叶片为试材介绍了免疫共沉淀的基本方法。主要试剂50uM雌激素，液氮，提取缓冲液TBS(50mM Tris-Cl，0.15M NaCl，pH 7.4，1mM PMSF，5mM Benzamidine)，glycine-HCl，TBS，洗脱缓冲液(TBS，0.5mg/mL 3
Restriction－Digest
Materials:Restriction enzymes of choice, such as BamH1 and EcoRIRestriction enzyme reaction buffer, such as MULTI-CORE (TM) (Promega)70 % Ethanol100 %
PCR－clean－up
Following PCR, you often want to get rid of the PCR primers and Taq polymerase before the next step. This is necessary for sequencing PCR products or
Disruption－by－Fusion－PCR
Disruption by Fusion PCRDavid Amberg and Ellen Beasley1) In separate PCR reactions, amplify the 5' and 3' ends of the gene of interest with p
琼脂糖凝胶电泳，做胶加EB目的
染色啊。你agarose胶电泳跑DNA，根据大小不同电泳速度不同，可是最后你需要看啊，怎么看呢？可见光下看不见DNA的。所以加EB也好，SYBRgreen也好，我们实验室用的genefinder也好，都是将DNA染色，用特殊的化学分子与DNA结合，并且在特殊波长的激发光下发射荧光，荧光强度与DNA含
Preparing－Lambda－DNA
Preparing Lambda DNA1- Coliphage lambda DNA is a widely used vector for recombinant DNA. The middle third of its 48,000 bp contains no genes required