InGelDigestionofProteinsSeparatedbyPolyacrylamideGelElectrophoresis
1. Excision of protein bands (spots) from polyacrylamide gelsRinse the gloves you use with water to avoid traces of dust in your sample.Rinse the gel with water.Excise spots with clean pipette tip (f 2 mm) cutting as close to the edge of the spot as possible (to reduce the volume of ''background'' gel)Transfer gel spot into a well of a 96-microtitre plate (Costar # 3363 + 3092)2. Reduction and alkylat......阅读全文
Western－Blot－with－Platelet－Protein
OUTLINEWestern blot is a wide used technique to identify a target protein/s for the certain antibody.PROTOCOLPrepare platelets.Lyse washed platelets (
UV－Shadowing
UV shadowing is a technique for visualizing nucleic acids separated on acrylamide/urea gels. The technique utilizes shortwave UV light (254 nm) and a
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
包涵体表达蛋白的纯化方法
Joseph SambrookPeter Maccallum Cancer Institute and The University of Melbourne, AustraliaDavid W. RussellUniversity of Texas Southwestern Medical Cen
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Phosphoamino acid analysis:Mark Kamps's method1. Label your protein with 32Pi. Then subject the protein to SDS polyacrylamide gel electrophoresis
Chromatin－Electrophoresis
Dr. William H. Heidcamp, Biology Department, Gustavus Adolphus College Exercise 10.4 - Chromatin Electrophoresis LEVEL II Materials 14 M Ure
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
Gel－Shift－Assay－Systems
ProtocolsDownloadprotocol183kbpdf?Abstract for Gel Shift Assay SystemsThe gel shift, or electrophoretic mobility shift, assay provides a simple and ra
DNA酶切及凝胶电泳（gel－electrophoresis）2
三、试剂 1、5×TBE电泳缓冲液：配方见第一章。 2、6×电泳载样缓冲液：0.25% 溴粉蓝，40%(w/v) 蔗糖水溶液，贮存于 4℃。 3、溴化乙锭(EB)溶液母液:将EB配制成10mg/ml，用铝箔或黑纸包裹容器，储于 室温即可。 第三节 操作步骤 一、
甲醛洋菜胶体电泳（formaldehydeagarose－gel－electrophoresis）
甲醛洋菜胶体电泳 （formaldehyde-agarose gel electrophoresis）甲醛是一种常用的RNA 变性剂。在进行甲醛洋菜胶体电泳分析时，必须先配制含有甲醛的洋菜胶体，RNA 也必须先以甲醛及formamide 进行变性处理，以确保其二度结构充分被打开。由于甲醛可能
DNA酶切及凝胶电泳（gel－electrophoresis）1
第一节 概 述 一. DNA的限制性内切酶酶切分析 限制性内切酶能特异地结合于一段被称为限制性酶识别序列的DNA序列之内或其附近的特异位点上，并切割双链DNA。它可分为三类:Ⅰ类和Ⅲ类酶在同一蛋白质分子中兼有切割和修饰(甲基化)作用且依赖于ATP的存在。Ⅰ类酶结合于识别位点并随机的切割识别位
琼脂糖凝胶电泳（agarose－gel－electrophoresis）介绍
主要试剂：核酸电泳缓冲液有三种，即Tris-硼酸(TBE)、Tris-乙酸(TAE)和Tris-磷酸(TPE).TBE与TPE缓冲容量高，DNA分离效果好，但TPE在DNA段回收时含磷酸盐浓度高，容易使DNA沉淀.TAE缓冲容量低，但价格较便宜，因而推荐选用TBE.缓冲液中的EDTA可螯合二价阳离子
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
Comparative－assessment－of－glycosylation－of－recombinant－human－...（二）
Experimental sectionChemicals and reagentsOne lot of PuregonR-HP of 50 IU/0.5mL and two lots of PuregonR-HP of 100 IU/0.5mL (rhFSH) (Organon, Oss, N
Assay－of－superoxide－dismutase－activity1
Assay of superoxide dismutase activity by combining electrophoresis and densitometryAbstract. A modified technique was developed to assay superoxide d
DNA指纹法
DNA Fingerprinting (David F. Betsch)the theory, procedures and applications · DNA Fingerprinting (Polyarylamide Gel) (Caltech)BAC DNA sample
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实验概要We provide a protocol for SDS-PAGE, Protein Blotting, Immuno-Detection.主要试剂1. 0.3 M TRIZMA® base (Product No. T1503), 20% methanol.2. 0.025 M TRIZ
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
Immunoblot－Protocol
This method was successful in our lab using prostate tissue and for our specific objectives. Investigators must be aware that they will need to tailor
Preparation－of－Stroma,－Thylakoid－Membrane,－and－Lumen－Fractions－from－...
Preparation of Stroma, Thylakoid Membrane, and Lumen Fractions from Arabidopsis thaliana Chloroplasts for Proteomic AnalysisFor many studies regarding
反向微柱的准备Preparation－of－ReversedPhase－Microcolumns
INTRODUCTIONOne versatile strategy for sample cleanup prior to MALDI-MS analysis uses microscale columns designed for direct sample elution onto the M
Western－杂交
Western 杂交（主要内容如下）Preparing of Protein LysatesWestern BlottingFar Western BlottingSemi Dry BlottingStripping MembranesTrouble Shooting and OthersPrepa
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
Capillary－Electrophoresis
Capillary electrophoresis is a very sensitive analytical technique. Sample components are separated within a fused silica capillary using one of sever
双向琼脂糖凝胶电泳（agarose－gel－electrophoresis）实验
【实验目的】了解和掌握双向电泳技术，并学习用它来研究与DNA 复制相关的问题。【实验原理】DNA 分子有线状的，还有一些非线状的，如复制叉和重组DNA 结构。双向琼脂糖凝胶电泳技术就是被人们开发用以研究一些非线状DNA 分子的。双向琼脂糖凝胶电泳技术(2-D gel)实际上可分为两类：中性/中性
琼脂糖凝胶电泳（agarose－gel－electrophoresis）检测DNA
原理： 琼脂糖是从海藻中提取出来的一种线状高聚物，可作为电泳支持物，适用于分离大小范围在0.2-50kb的DNA片段。DNA分子的迁移率与分子量的对数值成反比关系。观察其迁移距离，与标准DNA片段进行对照，就可获知该样品分子量大小。在质粒抽提过程中，由于各种因素的影响，使质粒DNA呈现超螺旋的共
DNA琼脂糖凝胶电泳（agarose－gel－electrophoresis）分析
一、原理琼脂糖凝胶具有分子筛效应。在中性ppH值的电泳缓冲液体系中，DNA分子由于带负电荷，所以在电场作用下由负极向正极泳动。由于DNA分子的大小和构型不同，在相同的时间内迁移至不同的位置。凝胶经溴化乙锭染色后，紫外检测仪下观察，即可看见DNA片段按大小不同呈条带分布。由于在一定条件下，DNA的迁移
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 sl
重组DNA的分离、克隆与测序实验手册7
III. Methods for DNA isolationA. Large scale double-stranded DNA isolationThe method used for the isolation of large scale cosmid and plasmid DNA is a
Two－Dimensional－Gel－Electrophoretic－Analysis－for－the－Human－Plasma－Proteome
OverviewThis protocol is a detail description of the laboratory procedure in performing 2D gel electrophoresis for illustrating the protein profile of