Polymerase Chain Reaction(PCR)One of the most powerful tools in molecular biologyInvented by Kary Mullis in 1983, resulting in his Nobel Prize in ChemistryIn essence, this process acts as a “copying machine” for DNAAdvances due to PCRGenerate very large quantities of specific DNA sequences, aiding in cloning Genomic sequencingDNA “fingerprinting”--forensic scienceDisea...

Software for:Making Restriction MapsDesigning PCR primersAssembling fragments, detecting SNPs, and managing raw sequence data For : GCG, Mac, PC, UNIX,    and on the Web  

Please contact Center for Plant Genomics (CPG) facility manager Hailing Jin (hljin@iastate.edu) regarding questions or corrections. REVERSE TRANSCRIPTION1. Add 1 ng in vitro transcribed RNA (human gene H2, GeneBank Accession #: AA418251) to the RNA sample (500 ng – 1000 ng)2. Add DEPC H2O to the RNA sample to 29.5 μl3. Add 0.5 μl 1μg/ul random hexamer and 1...

Angie L. Bookout and David J. MangelsdorfCorresponding Author: davo.mango@utsouthwestern.eduHoward Hughes Medical Institute, Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, TX, USA A major goal of the Nuclear Receptor Signaling Atlas (NURSA) is to elucidate the biochemical and physiological roles of nuclear receptors in vivo. Characteriz...

Introduction The RT-PCR kit contains polymerase and all necessary reagents to perform single tube RT and PCR. However, we will break the reaction into two discrete steps, RT (reverse transcription to convert mRNA to cDNA) and PCR (to amplify cDNA by the polymerase chain reaction). The RT reaction involves synthesizing cDNA from total RNA or polyA enriched RNA (mRNA). This c...

金红芝 杨志军摘 要：本文简述了NG和CT感染概况与分离培养法、涂片镜检法、酶免测定法、常规PCR法、实时PCR技术等主要的检测技术，着重介绍了实时PCR技术在NG和CT筛查中的应用情况。关键词：NG，CT，实时PCR，筛查

近年来，一类小分子核糖核酸（microRNA，也称miRNA）成为全球生命科学、生物医学、药物学等诸多领域关注的焦点。MicroRNA 是一种长度约18~23nt 的单链小分子RNA，由具有发夹结构且长度约70~90nt 的单链RNA 前体经过Dicer 酶切后生成。研究发现这些非编码的小分子RNA 绝大多数序列非常保守，它们的主要功能是调节生物体内相关的基因表达[1]。成熟的microRNA 一般只有二十几个碱基，同族的microRNA 之间通常...

基本步骤：1、目的基因（DNA和mRNA）的查找和比对；2、引物、探针的设计；3、引物探针的合成；4、反应体系的配制；5、反应条件的设定；6、反应体系和条件的优化；7、荧光曲线和数据分析；8、标准品的制备……………………

Real-time PCR 原理数学原理化学原理Real-time PCR 应用绝对定量相对定量等位基因分型阴阳性判定microRNA

张 蓓 ‘综 述 ， 沈 立 松 审 校(上海第二医科大学附属新华医院上海儿童医学中心实验诊断中心，上海200127)摘 要 : 多聚酶链反应飞速的发展使其成为了分子生物学实验室重要的工具，实时荧光定量PCR( real-timeq uantitative PCR)技术以其敏感性高、重复性好、速度快和污染少使PCR技术又向前迈进了一步。本文对real-time Q-PCR技术的原理、五种主要的荧光化学及定量方法作一介绍，同时也讨论了此技术...

什么是VentR、VentR（exo-）、 Deep VentR和 Deep VentR(exo-) DNA聚合酶?如何应用DNA聚合酶合成4－15 kb的PCR产物？为什么PCR反应后，没有产物或条带模糊？什么原因可以造成没有加入模板的阴性对照出现模糊条带？使用VentR或Deep VentR DNA聚合酶，产生什么样末端的DNA片段？用哪种克隆方法更好？PCR产物是否能在PCR反应体系中直接磷酸化？怎样才能提高PCR产物平滑末端的连接效率？VentR或Deep VentR DNA...