PolymeraseChainReaction(PCR)toAmplifyrRNAGeneFragment
Polymerase Chain Reaction (PCR) to Amplify rRNA Gene FragmentPrepare sufficient master mix for both partners (45 mL/50 mL reaction)10 mL 10x PCR buffer10 mL 2.5 mM dNTPs (0.25 mM final concentration)15 mL Primer A (5 pmole/mL)15 mL Primer B (5 pmole/mL)40 mL H2 O0.4 mL TaKaRa Ex-Taq DNA Polymerase (2 units)( Panvera )90 mLAliquot 45 mL of master mix into each of two 0.5 mL microfuge tube......阅读全文
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
Streptomyces:Protocols/PCR
Description Polymerase Chain Reaction (PCR) is a method of amplifying a specific DNA target sequence. The cycle involves denaturing the template doubl
利用Mastercycler－X50－PCR仪与epMotion系统实现高重复性的微...
利用Mastercycler X50 PCR仪与epMotion系统实现高重复性的微量体积PCR实验Highly reproducible low Volume PCR with Mastercycler® X50 and epMotion® Arora Phang, Tim Schommartz,
Cloning－PCR－products－using－TA－vectors
Cloning PCR products using TA vectorsby Paul N. Hengen, Ph.D. *Methods and reagents is a unique monthly column that highlights current discussions in
逆转录聚合酶链反应（RTPCR，rtpcr）
逆转录-聚合酶链反应 （Reverse Transcription-Polymerase Chain Reaction，RT-PCR）可以：(1）分析基因的转录产物；（2）获取目的基因；（3）合成cDNA探针；（4）构建RNA高效转录系统。实验方法原理逆转录-聚合酶链反应 (Reverse Tran
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
SemiQuantitative－RTPCR
The RT-PCR method can be used not only to detect specific mRNAs but also to semi-quantitate their levels. Thus, one can compare levels of transcripts
PCR的原理
聚合酶链式反应简称PCR（英文全称：Polymerase Chain Reaction）， 聚合酶链式反应简称PCR。聚合酶链式反应，其英文Polymerase Chain Reaction(PCR)是体外酶促合成特异DNA片段的一种方法,由高温变性、低温退火（复性）及适温延伸等几步反应组成一个周期
DNA标记
DNA标记（主要内容如下） DNA Labeling by Nick Translation Random Primed Labeling End-Labeling Purification of Labeled DNA Non-isotopic Labeling OthersDNA L
沙眼衣原体实验诊断技术进展
1907年，捷克学者Halberstaeder和Prowazek发现沙眼包涵体，1956年我国学者汤飞凡等分离沙眼衣原体成功，引起了全世界对其进行广泛深入的研究［1］。沙眼衣原体(chlamydia trachomatis, Ct)与人类疾病关系密切，且目前已成为许多国家和地区常见的性传播疾
Isolation－of－PBMCs－from－patient－blood－and－buffy－coats1
1| Transfer heparinized venous blood of patient to plastic50-ml tubes and dilute with an equal volume of PH buffer.When sodium citrate has been used a
MicroRNA－Expression－Profiling－by－Bead－Array－1
MicroRNA Expression Profiling by Bead Array Technology in Human Tumor Cell Lines Treated with Interferon-Alpha-2aMicroRNAs are positive and negative r
PCR实验指导与常见问题分析1
CONTENTPCR guide: a discussion of the main parameters influencing the outcome of the PCR and multiplex PCR reaction in 16 pages/sections and using ove
细菌感染的分子生物学检测技术
（一）常用的分子生物学技术1.核酸杂交 常用的杂交技术有：斑点杂交、southern印迹、原位杂交、Northern印迹等。探针的种类有全染色体DNA探针、染色体克隆片段DNA探针、质粒DNA探针、rRNA基因探针、寡核苷酸探针等医`学教育网搜集整理。2.核酸扩增技术 核酸扩增技术是分子生物学中最具
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
Competitive－RTPCR－Strategy－for－Quantitative－Evaluation－4
We have also been able to detect expression of this receptor in all studied tissues, which is consistent with the pleiotropic nature of growth hormone
Transcription,－Translation－of－S35Radiolabelled－Protein－and－Binding－to－GST
Prepare the template by linearizing 25ug plasmid DNA at the 3'' end of the insert. Phenol / chloroform extract, ethanol / NaCl precipitate and
How－do－you－synthesize－your－dsRNA
We routinely produce dsRNA by in vitro transcription of a PCR generated DNA template containing the T7 promoter sequence on both ends (I. Primer Desig
PCR－Primers－For－Gene－Expression－Detection－or－Quantification
Why PrimerBank?PrimerBank is a public resource for PCR primers. These primers are designed for gene expression detection or quantification (real-time
RNA－polymerase－III－transcription
Unlike the RNA polymerase II that transcribes a large variety of genes that encode proteins, RNA polymerase III transcribes only a limited number of g
RTPCR技术的定义
RT-PCR，反转录·聚合酶链反应（reverse transcription-polymerase chain reaction, RT-PCR）是将RNA的反转录（reverse transcription ）和cDNA的聚合酶链式扩增（PCR）相结合的技术。
PCR实验指导与常见问题分析2
Fig. 11. Example of the influence of extension temperature. Multiplex PCR with mixtrues A-B using two different PCR programs. Reactions on the right s
什么是PCR聚合酶链式反应－？
PCR即聚合酶链式反应 (Polymerase Chain Reaction，以下简称PCR)，是一种用于放大扩增特定的DNA片段的分子生物学技术，可理解为生物体外特殊的DNA复制。
DNA连接酶的用途
聚合酶链反应(polymerase chain reaction, PCR)。Taq酶扩增的PCR产物，3’末端总是带有1个非模板依赖型的突出碱基，而这个碱基几乎总是A，因为Taq酶对dATP具有优先聚合活性，故可用T载体克隆。
DNA聚合酶的用途
聚合酶链反应(polymerase chain reaction, PCR)。Taq酶扩增的PCR产物，3'末端总是带有1个非模板依赖型的突出碱基，而这个碱基几乎总是A，因为Taq酶对dATP具有优先聚合活性，故可用T载体克隆。
Competitive－RTPCR－Strategy－for－Quantitative－Evaluation－2
Determination of Accuracy of the Competitive PCRTo test the precision of the results obtained with this competitive PCR, five different amounts of T (
cDNA－Libraries
cDNA LibrariesIsolation of corresponding genetic informationInstead of synthesizing a desired gene, can we used the amino acid information to directly
PCR实验指导与常见问题分析4
Fig. 25. Multiplex PCR of mixtures A-D comparing PCR programs with 2 (green) and 1 (yellow) minute extension time at 54° C annealing temperature. Comp
Real－Time－PCR－Primer－Sets
Real Time PCR Primer SetsNOW OVER 300 PRIMER SETS!!!UPDATED: NOVEMBER 10th, 2003Quantitative RT-PCR is an important step for the validation of express
来自耶鲁大学的PCR常见问题的精辟总结
Troubleshooting discussion is based on the PCR protocol as described in the table below. All reactions are run for 30 cycles. COMPONENTVOLUMEFINAL CON