AnEconomicPCREnhancerforGCRichPCRTemplates

Since PCR is often problematic on GC-rich templates, we have evaluated different PCR enhancing additives and generated an Combinatorial Enhancer Solution (CES). We tested this solution on approx. 50 different primer pairs using several different polymerasesThe 5x concentrated Combinatorial Enhancer Solution (CES) contains:2.7 M betaine6.7 mM DTT6.7% DMSO and55 μg/ml BSAAdd this solution to your PCR and see if it work......阅读全文

An-Economic-PCR-Enhancer-for-GCRich-PCR-Templates

Since PCR is often problematic on GC-rich templates, we have evaluated different PCR enhancing additives and generated an Combinatorial Enhancer Solut

PCR-Additives

A variety of PCR additives and enhancing agents have been used to increase the yield, specificity and consistency of PCR reactions. Whilst these addit

PCR佐剂手册

A variety of PCR additives and enhancing agents have been used to increase the yield, specificity and consistency of PCR reactions. Whilst these addit

PCR进阶——GCRich片段的扩增策略

  PCR早已是分子生物学实验的常规技术,但是GC-rich扩增始终是有难度的应用。  以人基因组为例,大约3%的序列可划为GC-rich序列,尤其是在启动子,增强子等控制元件,GC-rich序列更为常见。因此,GC-rich片段的PCR扩增困难,也就阻碍了有关这类序列的科研进展。  GC-Rich

Cycle-Sequence-Reactions-For-Large-Insert-Plasmid-Templates

The following dye-labeled terminator reaction chemistries have been designed to balance conservation of reagents with the resulting sequence product s

利用Mastercycler-X50系列PCR独有的2D梯度实现高效的PCR条件...

利用Mastercycler  X50系列PCR独有的2D梯度实现高效的PCR条件优化Ultimate PCR Optimization with Eppendorf Mastercycler® X50 2D-gradientArora Phang, Tim Schommartz,Eppendorf

The-informationprocessing-pathway-at-the-IFNbeta-enhancer

The packaging of eukaryotic DNA into nucleosomes inhibits the access of factors to DNA and results in the repression of transcription, replication and

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

高GC含量PCR样本的扩增困难与防止污染与降解的操作方法

       每天做实验小心翼翼,时时担心提取的DNA被污染、被降解。终于,废了九牛二虎之力,好不容易提取到高质量的DNA,然而PCR仍然P不出来条带。是试剂加漏了吗?是酶失活了吗?换上新试剂,冰上重来一次,然并卵,不仅P不出来,更重要的是也不知道为啥P不出来,贝多芬都弹不出我的悲伤!    

如何分析DNA测序结果

Interpreting DNA Sequencing Results Evaluating ChromatogramsMany problems with sequencing results are not recognized by viewing the text file alone. T

定量RTPCR-(Quantitative-RTPCR)

Application: Quantitative RT-PCR is used to quantify mRNA in both relative and absolute terms. It can be applied for the quantification of mRNA expres

eRNA与Super-Enhancer-RNA在转录调控中扮演的角色

增强子是真核生物中关键的顺式作用基因调控元件,能有效地促进基因表达。它们可以通过作为转录因子和辅助因子的结合平台来维持转录的精确控制。超级增强子是由一簇典型增强子串联组成的具有更强转录调控能力的顺式元件。而全基因组分析发现增强子和超级增强子可以普遍进行转录,产生eRNA和SE-lncRNA。它们都具

Taq酶PCR实验方法介绍

General AdvicePCR allows the production of more than 10 million copies of a target DNA sequence from only a few molecules. The sensitivity of this tec

PfuDNA聚合酶PCR实验方法介绍

General AdvicePCR allows the production of more than 10 million copies of a target DNA sequence from only a few molecules. The sensitivity of this tec

长距离PCR

·         Long PCR (Church Lab)PCR conditioning for different templates, primer design, and moreLong PCR Reagents and Guidelines (Harusr/locald)Detail

高灵敏度化学发光技术应用心得(四)

结论:采用Bio-Rad的冷CCD化学发光成像系统进行化学发光的检测,比传统的暗室曝光方法更为简便,也大大缩短了实验时间。但在成像时需要注意选择合适的化学发光试剂,以适合CCD的成像检测。且大部分的化学发光试剂均对应暗室曝光的方式,因此在进行检测需要进行调整,如对发光液不能进行稀释,而必须要使用原液

LongPCR-Reagents-and-Guidelines

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

Long-PCR-Reagents-and-Guidelines

George Church Lab, Harvard Medical SchoolPCR_protocol.html">http://twod.med.harvard.edu/labgc/estep/longPCR_protocol.htmlEfficient Long PCR results fr

高GC比模板的PCR扩增

PCR条件的优化是一个麻烦耗时的过程,涉及到温度、时间、镁离子、引物、dNTP、Taq酶、模板等多个因素的调整。一般来说利用热启动,比如Platinum Taq DNA聚合酶(Invitrogen)可以达到更高的特异性,降低对镁离子浓度的依赖,并且有利于提高“问题模板”的产量。然而,传统的PCR条件

高GC比模板的PCR扩增

PCR条件的优化是一个麻烦耗时的过程,涉及到温度、时间、镁离子、引物、dNTP、Taq酶、模板等多个因素的调整。一般来说利用热启动,比如Platinum Taq DNA聚合酶(Invitrogen)可以达到更高的特异性,降低对镁离子浓度的依赖,并且有利于提高“问题模板”的产量。然而,传统的PCR条件

高GC比模板的PCR扩增

PCR条件的优化是一个麻烦耗时的过程,涉及到温度、时间、镁离子、引物、dNTP、Taq酶、模板等多个因素的调整。一般来说利用热启动,比如Platinum Taq DNA聚合酶(Invitrogen)可以达到更高的特异性,降低对镁离子浓度的依赖,并且有利于提高“问题模板”的产量。然而,传统的PCR条件

CRISPRCas9基因编辑的两种不同编辑实验流程的应用(三)

Alt-R® CRISPR-Cas9实例分析1、特别优化的crRNA/tracrRNA/sgRNA长度,提高编辑性能以HPRT基因中的12个位点为靶点,分别使用Alt-R® crRNA:tracrRNA、Alt-R® crRNA XT:tracrRNA、Alt-R® sgRNA,与Alt-R®

Protocol-for-dsRNA-Synthesis

实验概要        We routinely produce dsRNA by in vitro transcription of a PCR generated DNA template containing the T7 promoter sequence on both ends

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

DNA测序

DNA测序(主要内容如下)·         Sequencing Gel Preparation·         Preparation of Templates ·         DNA Sequencing by the Dideoxy Method·         DNA Sequen

SuperScript™-III-OneStep-RTPCR-System-with-Platinum®-Taq-High-Fidelity

实验概要The  SuperScript™ III One-Step RT-PCR System with Platinum® Taq  High  Fidelity is designed for sensitive, high-fidelity end-point detection  and

Long-PCR

Two long PCR steps:First round of the nested PCR step of the end point dilution procedure to quantitate the cDNA.First round of the nested PCR step of

Long-PCR

Two long PCR steps:First round of the nested PCR step of the end point dilution procedure to quantitate the cDNA.First round of the nested PCR step of

PCR实验指导与常见问题分析3

Influence of annealing temperature and number of loci amplifiedLike any other PCR, multiplex reactions should be done at a stringent enough temperatur

其它PCR方法

·         Standard PCR Protocol (Molecular Biology Techniques Manual)The followings are described in detailRecommended Reagent ConcentrationsRecommend