DNA实验技术

I did stupid thing after cloning a sequence into a plasmid just find out not in frame with the following sequence. just need insert one single nucleoacid will be ok. Any method to insert only one nucl ...

This is the URL to my gene of interest. I am trying to estimate the promoter region of this gene so that I can do Meth specific PCR on the promoter..the promoter region of this gene has not been posit ...

一、大肠杆菌质粒DNA的提取 质粒DNA的提取是从事基因工程工作中的一项基本实验技术，但提取方法有很多种，以下介绍一种最常用的方法： 碱裂解法：此方法适用于小量质粒DNA的提取，提取的质粒DNA可直接用于酶切、PCR扩增、银染序列分析。方法如下： 1、接1%含质粒的大肠杆菌细胞于2ml LB培养基。 2、37℃振荡培养过夜。 3、取1.5ml菌体于Ep管，以4000rpm离心3min，弃上清液。 ...

Donis-Keller Lab Manual Department of Genetics in Washington University School of Medicine http://hg.wustl.edu/hdk_lab_manual/plasmid/plsmid08.html Purpose: To utilize competent E. coli bacteria to r ...

真核细胞的mRNA在加工过程中有一个比喻为“穿鞋戴帽”的过程，因此mRNA的3'末端都带有一段Poly A，这是利用逆转录酶制备cDNA文库的基础。但是由于cDNA的5'端的序列各不相同，如何获得全长的cDNA，如何扩增由微量的mRNA逆转录得到的cDNA文库、如何利用已知片断序列得到全长的cDNA（即RACE），曾经是一个令人困扰的问题。 常见的做法是在合成cDNA的双链 ...

Hancock Laboratory Methods. Department of Microbiology and Immunology University of British Columbia British Columbia Canada http://www.cmdr.ubc.ca/bobh/showmethod.php?methodid=67 METHOD: 1.Run gel a ...

概 述 基因组DNA的提取通常用于构建基因组文库、Southern杂交(包括RFLP)及PCR分离基因等。利用基因组DNA较长的特性，可以将其与细胞器或质粒等小分子DNA分离。加入一定量的异丙醇或乙醇，基因组的大分子DNA即沉淀形成纤维状絮团飘浮其中，可用玻棒将其取出，而小分子DNA则只形成颗粒状沉淀附于壁上及底部，从而达到提取的目的。在提取过程中，染色体会发生机械断裂，产生大小不同的片段，因此 ...

I am working with very small amount human tissue. Is there anyone who had the experience of using glycogen with 100% ethanol in pheno/chloroform DNA extraction protocol ? Does it realy work to increas ...

质粒是存在于细菌染色体外的一个或多个能独立复制并稳定遗传的小型环状双链DNA分子，其分子量一般在0. 2～10kD 范围内。由于质粒分子小，便于分离和提取，可以携带目的基因进入细菌、动物细胞或植物体内进行扩增与表达。基因克隆实验中常将经改造后的质粒作为运送基因的载体，质粒D2NA提取效率及质量对后续实验步骤(如酶切、连接、转化大肠杆菌与PCR扩增等)的成功与否有着直接的关系，因而高效快速地从细菌细 ...

Materials: Silica Suspension: add 2 g of silica to 15 ml of H2O wash 3x by centifugation at 2000 x g for 2 min estimate vol of silica and resuspend in 2 vol H2O Silica Wash Solution: 50 mM NaCl 10 mM ...

Hi can anyone answer this I am having problems with my southern hybridization. I am wondering if the DNA transfers to my membrane. Is it possible that the DNA can wash into the solutions when denaturi ...

根据载体的遗传特征筛选重组子，如α-互补、抗生素基因等。现在使用的许多载体都带有一个大肠杆菌的DNA的短区段，其中有β-半乳糖苷酶基因（lacZ）的调控序列和前146个氨基酸的编码信息。在这个编码区中插入了一个多克隆位点（MCS），它并不破坏读框，但可使少数几个氨基酸插入到β-半乳糖苷酶的氨基端而不影响功能，这种载体适用于可编码β-半乳糖苷酶C端部分序列的 ...

Tissue collection storage microdissection sectionin g: See separate protocol. Tissue handling : Note that all fresh tissue shoμld be handled as BioSafety Level 2 materials (wear gloves lab coat etc ...

摘　要　对植物中DNA抽提是在分子水平上对植物进行系统分析与研究的基础性工作.经多次实验，使用改良的CTAB法抽提山茶属植物中总DNA，即在抽提前加入抗氧化剂β-巯基乙醇，并使用冷冻的异丙醇来沉淀DNA， 获得了较好的效果，其纯度和得率完全能满足常规分子生物学操作的要求. 关键词　山茶属植物；DNA抽提；叶片；改良CTAB法 中图法分类号　Q523；Q949.758.4 DNA Ex ...

(adapted from Bruce A. Roe Department of Chemistry and Biochemistry The University of Oklahoma Norman Oklahoma 73019 broe@ou.edu) Typically 2.5 - 3 volumes of an ethanol/acetate solution is added to t ...

Hi all! We are trying to extract genomic DNA from the leaves of an Ericacea (we suppose it is rich in tanins and phenolics) and we can't do it! We have used two different kits which work OK for other ...

Contributed by Dr. A. Gratchev Single Nucleotide Primer Extension is a powerful method which can be used for the precise analysis of methylation in a certain position. The procedure is shown on the fi ...

1.Vectorpedia： 输入质粒骨架，直接查询，非常方便; http://www.addgene.org/pgvec1?f=v&cmd=listvecinfo 2. google 检索式子：质粒名 ＋ map 质粒名 ＋sequence ＋filetype:txt or filetype:pdf 或者：进入google，点击"图片搜索"，直接查找图谱。 3.go ...

Does anyone have expereince of extracting DNA from urine? I am having problems with this. I have previously extracted the DNA but then it degraded very quickly and since then I havent even been able t ...

近年来抗性基因研究的突破性进展、抗性基因的克隆和序列分析所揭示的其编码蛋白的组成、拓扑学和亚细胞定位等特征为揭开抗性基因的作用特点提供了线索。一般来讲基因克隆的策略可分为两种:正向遗传学途径和反向遗传学途径。前者以欲克隆的基因所表现的功能为基础通过鉴定其产物或某种表型的突变进行如功能克隆( Functional Cloning) 和表型克隆( Phenotype Cloning) ;后者则着眼于基 ...