EthidiumBromideDecontamination
N.B.: Ethidium bromide is a powerful mutagen. Protective gloves should be worn at all times when handling solutions containing ethidium bromide.Decontamination of solutions > 0.5mg/mlThis method reduces the mutagenic activity by ~3000 fold and is taken from Quillardet and Hofnung, 1988.1) Add sufficient water to make the concentration < 0.5mg/ml.2) Add 1 volume of 0.5M KMnO4 followed by one volume of ......阅读全文
琼脂糖凝胶电泳检测DNA
目的:学习水平式琼脂糖凝胶电泳,检测质粒DNA的构型、分子量的大小。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。
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
Genomic-Southern-Blot
SolutionsProtocol:Digest 5-10 μg genomic DNA overnight with restriction enzyme of choice.Run digested gDNA on 0.8% TAE gel with marker (with no ethidi
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
微生物拟核体内体外染色观察实验_溴化乙锭染色法
实验方法原理如果将微生物细胞裂解,使其拟核(即染色体 DNA)被抽提出来,通过溴化乙淀(ethidium bromide,简称 EB)染色并进行琼脂糖凝胶电泳,便可观察到释放到细胞外的染色体 DNA。EB 是一种扁平分子染料。可特异性插入 DNA 碱基对之间,在紫外线照射下,使 DNA 呈现荧光,因
Protocol-to-Count-Cell-Number-of-Preimplantation-Embryos
Protocol to Count Cell Number of Preimplantation Embryos using Nuclear Staining with Hoechst 33342 or DAPI Introduction The following is a simple pro
DAPI染色液使用说明
DAPI染色液(DAPI Staining Solution)是经过精心优化几乎适用于所有常见细胞和组织细胞核染色的染色液。 DAPI,即2-(4-Amidinophenyl)-6-indolecarbamidine dihydrochloride,也称DAPI dihydrochlor
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
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
Nested-RTPCR-for-Hepatitis-C-from-Paraffin-Sections
RNA Extraction from Histologic SectionsUnstained 4 µm thick sections of formalin fixed paraffin embedded liver biopsies were transferred from glass sl
实验室常用英文术语集锦(三)
培养箱incubator 超滤器Ultrahigh Purity Filter 超低温冰箱Ultra-low Temperature Freezer 超声破碎仪Ultrasonic Cell Disruptor 紫外观察灯Ultraviolet Lamp
Native-chromatin-immunoprecipitation-protocol
实验概要Native chromatin immunoprecipitation to query specific chromatin states of individual genes. 主要试剂10 x TBS 0.1 M Tris-HCl (pH 7.5) 1.5 M NaCl 30 mM
自然染色质免疫沉淀实验设计
1. The preparation of native chromatin from cultured human cells1.1.Cultured cells (e.g. HL-60 or lymphoblastoids) are grown to a density of approxima
Native-chromatin-immunoprecipitation-protocol
实验概要The method is a native chromatin immunoprecipitation protocol.主要试剂1. 10 x TBS 0.1 M Tris-HCl (pH 7.5) 1.5 M NaCl 30 mM CaCl2 20 mM MgCl2 50 mM Na
关于维库溴铵的基本信息介绍
1、基本信息 中文名称:维库溴铵 中文别名:溴化凡寇罗宁; 英文名称:vecuronium bromide 英文别名:Vecuronium bromide; Vecuronium Bromide; Vecuronium Bromide Hydrate; CAS号:50700-72-6
哺乳动物RNAi技术-Mammalian-RNA-Interference
Mammalian RNA InterferenceThomas TuschlLaboratory for RNA Molecular BiologyThe Rockefeller University, New York Excerpted from RNAi: A Guide To Gene S
Guide-to-Cell-Proliferation-and-Apoptosis-Methods
Chapter 1: Cell Death - Apoptosis and Necrosis1.1Introduction21.1.1Terminology of cell death21.1.2Differences between necrosis and apoptosis31.1.3Apop
EB和紫外辐射对实验人员的危害需引起注意
在电泳实验或其他核酸分离实验中,常用EB(Ethidium bromide,EtBr)作非放射性的Marker,来识别和显示核酸条带。EB是一种深红色不易挥发的晶体,微溶于水。当被紫外线照射时能发出红褐色的荧光。尽管EB是一种高效的显色剂,但它的高危险性还是需要引起您的特别注意。EB是一种强诱变
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
关于泮库溴铵的物化性质介绍
一、泮库溴铵的基本信息: 中文名称:泮库溴铵 中文别名:巴活朗 英文名称:pancuronium bromide 英文别名:Pancuroniumdibromide; Pancuronium bromide; Pancuronium Bromide; CAS号:15500-66-0
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
Bench-Top-Radioactive-Work-Protocol
Log in radioactive material received and deduct amount of radioactive material used in each experiment in Radioisotope Log Book.Clear your bench top w
简述奥替溴铵的物化性质
一、基本信息 中文名称:奥替溴铵 中文别名:异雄酮.异雄酮;N,N-二乙基-N-甲基-2-[[4[[2-(辛基)苯甲酰]氨基]苯甲酰]氧基]乙烷铵溴化物;奥替溴胺 英文名称:Octylonium Bromide 英文别名:Otilonium Bromide; Ethanaminium (
凝胶成像系统内主要光源的介绍
凝胶成像系统内主要光源的介绍,凝胶成像系统,顾名思义以一套可对凝胶进行成像的光学系统,而光源便是其光路系统的重要组成部分,可以说没了光源,成像系统几乎无法运行使用,但与CCD、镜头以及分析软件相比,往往却没有得到应有的重视。我们经常遇到这样的用户,他们知道系统光源出现故障或整个系统报废的时候都不知道
凝胶成像系统内主要光源的介绍
凝胶成像系统内主要光源的介绍,凝胶成像系统,顾名思义以一套可对凝胶进行成像的光学系统,而光源便是其光路系统的重要组成部分,可以说没了光源,成像系统几乎无法运行使用,但与CCD、镜头以及分析软件相比,往往却没有得到应有的重视。我们经常遇到这样的用户,他们知道系统光源出现故障或整个系统报废的时候都不
“质粒DNA超速离心分离”的补充说明
一)用CSCL-E.B.梯度分离大肠杆菌中质粒DNA、染色体DNA、蛋白质及RNA的详细步骤(1) 溶液制备:溶液Ⅰ:50mM葡萄糖,10mM EDTA,25 mM Tris-HCL (PH8.0)。溶液Ⅱ:0.2M NaOH,1%SDS。溶液Ⅲ:3M醋酸钠(PH4.8)TE缓冲液:10 mM Tr
PCR-各种缓冲液的配方
PCR 各种缓冲液的配方电泳缓冲液50×Tris-乙酸(TAE)缓冲液 终浓度配制1L溶液成分 各成分的用量2mol/L Tris碱 242g1mo
电泳缓冲液、染料和凝胶加样液的配置
电泳缓冲液50×Tris-乙酸(TAE)缓冲液成分及终浓度 配制1L溶液各成分的用量 2mol/L Tris碱1mol/L 乙酸100 mmol/L EDTA水 242g57.1 ml的冰乙酸(17.4 mol/L)200ml的0.5 mol/L EDTA(pH 8.0)补足1L 5×Tris-硼酸
Preparation-of-Plasmid-DNA-by-Alkaline-Lysis-with-SDS:-Maxipreparation
实验概要Plasmid DNA is isolated from large-scale (500 ml) bacterial cultures by treatment with alkali and SDS.主要试剂Buffers and SolutionsAlkaline lysis solu
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