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
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
多功能紫外分析仪使用技巧
多功能紫外分析仪使用技巧一、使用范围介绍:ZF1-II多功能紫外分析仪系,又称、切胶仪、采用不同波长的紫外光对DNA、RNA电泳凝胶样品进行观察拍照、检测蛋白质、核甘酸、适用于核酸电泳分析、检测,PCR产物检测,DNA指纹图谱分析,纸层分析或薄层分析等。紫外分析仪适用于核酸电泳、荧光的分析、检测,
基本实验技术
I. Safety ProceduresA. ChemicalsA number of chemicals used in this laboratory are hazardous. All manufacturers of hazardous materials are required by
琼脂糖凝胶中DNA的检测
琼脂糖凝胶中的核酸通过染色,可以在波长为 300 nm 的紫外灯下检测。介绍琼脂糖凝胶中核酸染色的两种方法:溴化乙锭(ethidium bromide, EB ) 染色法和 SYBR Gold 染色法。本实验来源「分子克隆实验指南第三版」黄培堂等译。实验方法原理琼脂糖凝胶中的核酸通过染色,可以在波长
DNA酶切及凝胶电泳的相关内容介绍
琼脂糖是从海藻中提取出来的一种多聚多糖,是由D-半乳糖和L-半乳糖以α-1,3和β-1,4糖苷键相连形成的线状高聚物。琼脂糖遇冷水膨胀,溶于热水成溶胶,冷却后成为孔径范围从50nm到大于200nm的凝胶。 琼脂糖或聚丙烯酰胺凝胶电泳是分离鉴定和纯化DNA片段的标准方法。该技术操作简便快速,可以
DNA拓扑异构酶催化反应的相关介绍
很多,其反应本质是先切断DNA的磷酸二脂键,改变DNA的链环数之后再连接之,兼具DNA内切酶和DNA连接酶的功能.然而它们并不能连接事先已经存在的断裂DNA,也就是说,其断裂反应与连接反应是相互耦联的。拓扑异构酶(包括Ⅰ型和II型)都可以用符号转化模型进行解释 除了DNA拓扑异构酶可以产生异构
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
Genotyping-Transgenic-Rodents-by-PCR
Genotyping Transgenic Rodents by PCRThis is how we test mice and rats for the presence of the transgene by PCR. It is provided for those investigators
DNA转化实验指导3
6. Simultaneous digestion of the pUC vector with both enzymes in the presence of 3 units of Shrimp Alkaline Phosphatase (Amersham BioSciences) in
PCR产物纯化方法
Purification of PCR Products in Preparation for CloningJoseph SambrookPeter Maccallum Cancer Institute and The University of Melbourne, AustraliaDavid
Methylation-Specific-PCR
Methylation Specific PCRProtocol written by James Herman*Methylation Specific PCR (MSP) is a simple rapid and inexpensive method to determine the meth
叶绿体DNA分离
设备:Hitachi CS-150GXL或CS-120GXL微量超速离心机,S100AT6 转头,5PA 密封管(如果用4PC管,可接比例减少各层液量)溶液配制:A液:0.35Msorbitol(山梨醇),50mM Tris—Hcl (PH8.0) 25mM EDTA—Na2B液:5%(w/w)So
PCR-protocol
PCR reactionProtocol for 50µl reaction - adjust amounts if necessary, for a 20µl reaction use the same volumes of primer and dNTP-mix, but adjust the
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
EB染色液使用说明
产品简介:EB染色液是EB的水溶液,浓度为10mg/ml。EB(溴化乙锭)能与核酸分子特异结合,用于观察琼脂糖或聚丙烯酰胺凝胶中DNA或RNA分子。广泛应用于核酸电泳前后的染色以及流式细胞仪的样品染色处理等。溴化乙锭(Ethidium Bromide;C21H20N3Br;分子量为394.32)含有
核酸电泳的指示剂与染色剂
指示剂 核酸电泳常用的指示剂有溴酚兰和二甲苯青及银染色.溴酚兰在碱性液体中 呈紫兰色,在0.6%、1%、1.4%和2%琼脂糖凝胶电泳中,溴酚兰的迁移率分别与1Kb、 0.6Kb、0.2Kb和0.15Kb的双链线性DNA片段大致相同.二甲苯青的水溶液呈兰色,它在 1%和1.4%琼脂糖中电泳
核酸电泳的指示剂与染色剂
指示剂 核酸电泳常用的指示剂有溴酚兰和二甲苯青及银染色.溴酚兰在碱性液体中 呈紫兰色,在0.6%、1%、1.4%和2%琼脂糖凝胶电泳中,溴酚兰的迁移率分别与1Kb、 0.6Kb、0.2Kb和0.15Kb的双链线性DNA片段大致相同.二甲苯青的水溶液呈兰色,它在 1%和1.4%琼脂糖中电泳时,
Electrophoresis-of-PCR-products-with-Sunrise-gel-apparatus
Electrophoresis of PCR products with Life Technologies Sunrise gel apparatusGel: In a 500 ml Pyrex® glass bottle, add:Agarose:3 gH2O270 mls10X TA30 ml
核酸电泳指示剂和染色剂的选择
指示剂:核酸电泳常用的指示剂有溴酚兰和二甲苯青及银染色.溴酚兰在碱性液体中 呈紫兰色,在0.6%、1%、1.4%和2%琼脂糖凝胶电泳中,溴酚兰的迁移率分别与1Kb、 0.6Kb、0.2Kb和0.15Kb的双链线性DNA段大致相同.二甲苯青的水溶液呈兰色,它在 1%和1.4%琼脂糖中电泳时
过氧化氢蒸汽-(-H-P-V-)-生物有效性(二)
2. 参考文献/ 出处1. Rogers, J. V., C. L. Sabourin, Y. W. Choi, W. R. Richter, D. C. Rudnicki, K. B. Riggs, M. L. Taylor, and J. Ch a ng.2005. Decontaminat
Quick-and-reliable-method-to-analyze-meiotic-segregation-patterns
It is well known that multiple auxotrophic markers impede fruiting in Coprinus cinereus. Restriction fragment length polymorphisms have been used to a
RNA-analysis-on-nondenaturing-agarose-gel-electrophoresis
实验概要RNA analysis on non-denaturing agarose gel electrophoresis实验步骤1. The following gel electrophoresis conditions are recommended:- use 1X TAE buffer
DAPI染色液的使用方法及注意事项
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 dihydrochloride,分子式
RNA-analysis-on-nondenaturing-agarose-gel-electrophoresis
1. The following gel electrophoresis conditions are recommended:- use 1X TAE buffer instead of 1X TBE- use agarose gel in the concentration of 1.1%-1.
核酸的染色介绍
核酸染色法一般可将凝胶先用三氯乙酸、甲酸—乙酸混合液、氯化高汞、乙酸、乙酸镧等固定,或者将有关染料与上述溶液配在一起,同时固定与染色。有的染色液同时染DNA及RNA,如stains-all、溴乙锭、掊花青-铬矾法等,也有RNA、DNA各自特殊的染色法。1.RNA染色法(1)荧光染料溴乙锭(ethid
Protocol-for-competitive-RTPCR
For quantifying mRNA, we use a competitive RT-PCR protocol with internal standard RNAs. These are added in a defined quantity to the RNA sample prior
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
Competitive-RTPCR-Strategy-for-Quantitative-Evaluation-1
Competitive RT-PCR Strategy for Quantitative Evaluation of the Expression of Tilapia (Oreochromis niloticus) Growth Hormone Receptor Type IQuantizatio
DNA的纯化浓缩与定量
实验目的 将复杂的细胞分子混合物加入有机溶媒萃取以除去蛋白质及其它成分,就可以纯化DNA。一般常用酚及氯仿(phenol/chloroform)可使蛋白质变性(denaturaion)的特性来进行萃取的步骤,DNA和RNA不溶于有机溶媒中,而溶于水层。另外,分子选殖(molecular clon
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。