DNA基础知识

After amplification with a proofreading polymerase place samples on ice and add 0.7-1 unit of Taq polymerase per tube directly into the PCR reaction tube. Mix well. Incubate at 72°C for 8-10 minutes. ...

This protocol uses Promega''s pGEM-T kit (#A3600).PCRFor TA cloning it is optimal if the PCR primers have G''s at the 5'' end as this will maximize the probability of Taq polymerase adding the termina ...

Here is some informal background information that will hopefully provide support and context if you want to use ET recombination. Perhaps needless to mention this information has not been refereed is ...

AbstractFunctional genomics require manipulation and modification of large fragments of the genome. Such manipulation has only recently become more efficient due to the discovery of different techniqu ...

A stab culture is made by inoculating bacteria into a vial containing LB agar with the appropriate antibiotic. After overnight incubation bacterial growth should be visible both in the puncture and on ...

Proteinase K digestion Mix DNA extraction buffer 98 μl ReagentB 2 μl ProteinaseK Mix fresh. 100 μl is enough for a small pea size chunk of tissue or one embryo Place small piece of tissue or embryo in ...

Material100% ethanol (for analysis) sodium acetate -20°C freezer Optional: you can use linear polyacrylamide glycogen or tRNA as precipitation carrier --zurdo 03:45 9 October 2009 (EDT) tabletop centr ...

PREPARE SOLUTIONS 1. 10X Dephosphorylation buffer:0.5 M Tris-HCl pH 8.5 1 mM EDTA PROCEDURENOTE: 1 unit of Alkaline Phosphatase (AP) can hydrolyze 50 pmol of 5'' terminal phosphorylated DNA fragment ...

INTRODUCTION High-throughput whole-genome analysis has become a practical and important technique to understand nuclear processes such as transcription replication and genome structure. Though microar ...

操作步骤：（实验前请先阅读注意事项）提示：第一次使用前请先在漂洗液RW瓶加入指定量无水乙醇!操作前在裂解液RLT中加入β-巯基乙醇至终浓度1%，如1 ml RLT 中加入10μl β-巯基乙醇。此裂解液最好现用现配。配好的RLT 4℃可放置一个月。1.直接研磨法（推荐）:a. 新鲜植物组织称重后取100mg迅速剪成小块放入研钵（冰冻保存或者液氮保存样品可直接称重后取100mg放入研钵 ...

DL5000 DNA Marker.doc

The protocol for LIC by Exonuclease III梁耀极1. Design the primers with 15-bp overlap;2. Digest the vector by proper restriction enzyme;For getting high quality vectors there are some good advices as fol ...

PEI 转染法材料：质粒DNA指数生长的真核细胞PEI （聚乙烯亚胺）1×HBS （pH7.4）配方：PEI 储存液（100 μM）：称取125 mg PEI 粉末溶解于50 ml 1×HBS （pH7.4）中， 0.2 μm 滤膜过滤，储存于4℃备用。1×HBS （pH7.4）： 将8.76 g NaCl 溶解于900 ml 超纯水，加入20 ml 1 M 的HEPES，调pH 值到7.4，定容 ...

DNA以核蛋白形式存在于细胞核中，制备DNA的原则是既要将DNA与蛋白质、脂类和糖类等分离，又要保持DNA分子的完整。蛋白酶K及链霉蛋白酶E的应用使这两个原则得到了保证。蛋白酶K或链霉蛋白酶E均为广谱蛋白酶，其重要特性是能在SDS和EDTA存在下保持很高的活性。在匀浆后提取DNA的反应体系中，SDS可破坏细咆膜、核膜，并使组织蛋白与 DNA分子分离，EDTA抑制细胞中DNase活性，蛋白酶K或链霉 ...

（一）原理DNA 在酸性条件下加热，其嘌呤碱与脱氧核糖间的糖苷键断裂，生成嘌呤碱、脱氧核糖和脱氧嘧啶核苷酸，而2-脱氧核糖在酸性环境中加热脱水生成ω-羟基-γ-酮基戊糖，与二苯胺试剂反应生成蓝色物质，在595nm 波长处有最大吸收。DNA 在40-400μg 范围内，光吸收与DNA的浓度成正比。在反应液中加入少量乙醛，可以提高反应灵敏度。（二）试剂及器材1. DNA标准溶液准确称取小牛胸腺的DN ...

一、原理： 在浓氯化钠（1�2mol/L）溶液中，脱氧核糖核蛋白的溶解度很大，核糖核蛋白的溶解度很小，在稀氯化钠（0.14 mol/L）溶液中，脱氧核糖核蛋白的溶解度很小，核糖核蛋白的溶解度很大。因此，可利用不同浓度的氯化钠溶液，将脱氧核糖核蛋白和核糖核蛋白从样品中分别抽提出来。将抽提得的核蛋白用SDS（十二烷基硫酸钠）处理，DNA或（RNA）即与蛋白质分开，可用氯仿� ...

Methylation Specific PCRProtocol written by James HermaundefinedMethylation Specific PCR (MSP) is a simple rapid and inexpensive method to determine the methylation status of CpG islands. This approach all ...

Protocols Downloadprotocol183kb pdf?Abstract for Gel Shift Assay SystemsThe gel shift or electrophoretic mobility shift assay provides a simple and rapid method for detecting DNA-binding proteins. Thi ...

分子克隆常用载体　　DNA片段的克隆需要合适的载体，载体或是质粒，或是噬菌体，或是病毒，通常大多经过人工改造一、质粒常用的有pBR322，pUC系列质粒等。（一）pBR322质粒是4362bp的环状双链DNA载体，有2个抗药性基因（四环素和氨苄青霉素），一个复制起始点和多个用于克隆的限制酶切点。当缺失抗药性基因的大肠杆菌被pBR322成功地转化时，它便从该质粒获得了抗生素抗性。两个抗生素基因中均含 ...

Scanning the human genome with combinatorial transcription factor librariesPublished online: 18 February 2003 doi:10.1038/nbt794March 2003 Volume 21 Number 3 pp269-274Pilar Blancafort Laurent Magne ...