PCR引物设计

Detecting DNA Contamination in RT-PCR

Avoiding DNA Contamination in RT-PCR A frequent cause of concern among investigators performing quantitative RT-PCR is inaccurate data due to DNA contamination in RNA preparations. Although DNA contam ...

Designing PCR programsBasic Principles (see also Page 01) The requirement of an optimal PCR reaction is to amplify a specific locus without any unspecific by-products. Therefore annealing needs to tak ...

Single Primer ("Semi-Random") PCRJuly 26 2000 ECKDescriptionSingle primer PCR allows amplification from known to unknown regions in chromosomes phage plasmids large PCR products and other sources of D ...

This procedure was first described by Bertrand et al (Proceedings of the National Academy of Science USA (1993) Vol. 90 pp. 3496-3500 and Nucleic Acids Research (1994) Vol. 22 pp. 293-300) to demonstr ...

Protocol for competitive RT-PCR 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 to the RT reaction. ...

Successful RT-PCR requires a high quality intact RNA template. Use the following guidelines to help prepare this template: To minimize the activity of RNases that are released during cell lysis inclu ...

RT-PCR AnalysisSolutions10X RT Buffer10X PCR Buffer100 mM Tris pH 9.0500 mM KCl1% Triton X-10025 mM MgCl2use at a concentration of 1.5 mMLysis Solution4M GuSCN 250 g guanidine thiocyanate25mM Na citr ...

Question 1.What are the differences among RNase H RNase A RNase B and RNase C?2.In your cDNA kits RNase H is added in the second strand reaction to produce more nicked RNA as primers for DNA synthes ...

Question What is the highest temperature that SUPERSCRIPT II MMLV or THERMOSCRIPT can be used? Answer For RT-PCR SUPERSCRIPT II RT can be used up to 50 C. MMLV can be used at temperatures up to ...

Polymerase Chain Reaction (PCR) cont.Choice of Polymerases for PCROne of the important advances which allowed development of PCR was the availability of thermostable polymerases. This allowed initiall ...

The following colony PCR protocol has been designed to be performed in individual reaction tubes. We usually test three colonies from each transformation along with a wild-type control. A series of f ...

Site-directed Mutagenesis using PCRMichael P. Weiner Tim Gackstetter Gina L. Costa John C. Bauer and Keith A. KretzFrom: Molecular Biology: Current Innovations and Future Trends. Eds. A.M. Griffin and ...

INTRODUCTION The products of a PCR reaction - especially when this is done on eukaryotic genomic DNA and when using degenerate primers - often contain a mixture of discrete-sized bands one of which i ...

Cloning PCR products using TA vectorsby Paul N. Hengen Ph.D. *Methods and reagents is a unique monthly column that highlights current discussions in the newsgroup bionet.molbio.methds-reagnts availabl ...

PEG PRECIPITATION OF PCR PRODUCTSThis protocol can be used instead of EXO/SAP for removing excess primers and nucleotides from PCR products before cycle-sequencing. ...

Long-PCR Reagents and Guidelines from George Church as Modified from Cheng et al. (1) General Guidelines for Long-PCR Conditions and Enzyme Mixtures==========Following the results of Cheng et al. (1) ...

PCR技术简介 　前言 一滴残留在裙子上的精液使得美国总统Bill Clinton不得不坦承他与白宫实习生有不正当的关系。因为他知道现在的生物科技就连一个精子也能被用来做为证据。这种将极微量的生物标本化为可供鉴定的现代技术正是PCR(Polymerase chain reaction)--聚合酶链式反应具有的特色之一。这也是分子生物医学令人震撼的一例。 何谓PCR 简单的说，PCR就是利用DNA聚 ...

关键词：寡核苷酸；优化设计中图分类号：Q524 在核酸分子杂交、DNA序列测定和通过PCR放大DNA片段等实验中，都需要使用寡核苷酸作为探针或引物，而对这些反应的质量起最重要影响作用的，就是这些寡核苷酸探针或引物。用优化的寡核苷酸进行实验能够很快得到好的结果，而用不够合适的寡核苷酸时，常常得出似是而非的结果，不仅大大增加了后续实验的工作量，还可能一无所获。 怎样优化设计寡核苷酸呢?至 ...

聚合酶链反应一单链构象多态性分析 (PCR-SSCP) 　 聚合酶链反应-单链构象多态性分析（Single Strand Conformation Polymorphism Analysis of Polymerase Chain Reaction Products PCR-SSCP）是近年来发展起来的一种基因分析方法。 　PCR-SSCP分析的基本程序为：首先PCR扩增特定靶序列，然后将扩 ...