PlasmidorCosmidDNAMiniprep
This protocol can be used to isolate sufficient amount DNA from 1.5ml o/n culture or 3ml 6hr culture to do several enzyme digestions.Spin 1.5ml o/n culture at 1,2000rpm for 30 Sec. Discard the supernatant.Resuspend the pellet by vertex in 100ul QP buffer, R.T. 5min.Mix 200ul 10N NaOH and 500ul 20% SDS add water to final volume 10ml.Add 200ul the solution above to the suspension (#2), invert 5 times, R.T. 5min.Add 150......阅读全文
质粒是什么
质粒（plasmid）是细菌、酵母菌和放线菌等生物中染色体（或拟核）以外的DNA分子，存在于细胞质中，具有自主复制能力，使其在子代细胞中也能保持恒定的拷贝数，并表达所携带的遗传信息，是闭合环状的双链DNA分子。质粒不是细菌生长繁殖所必需的物质，可自行丢失或人工处理而消除，如高温、紫外线等。质粒携带的
质粒是什么
质粒（plasmid）是细菌、酵母菌和放线菌等生物中染色体（或拟核）以外的DNA分子，存在于细胞质中，具有自主复制能力，使其在子代细胞中也能保持恒定的拷贝数，并表达所携带的遗传信息，是闭合环状的双链DNA分子。质粒不是细菌生长繁殖所必需的物质，可自行丢失或人工处理而消除，如高温、紫外线等。质粒携带的
Electrotransformation－of－Lactobacillus－Spp.
OverviewGeneral guidelines for the electro-transformation of Lactobacillus sake as described by Berthier et al. and as used by Alegre et al. with Lact
通过细胞受体代谢生物素化进行图像分析
Metabolic biotinylation of mammalian cell receptors for imagingBakhos A. Tannous , btannous@hms.harvard.edu, Massachusetts General Hospital and Harvar
DNA疫苗实验方法
For direct gene transfer of tibialis anterior (TA) muscle in mice: It is optimal to use 6-8 week old mice (weight 19-21 gm). Females give better immun
酵母质粒载体的基本信息介绍
酵母质粒载体是基因表达载体的一种,既可以在大肠杆菌中、又可以在酵母系统中进行复制与扩增,所以也称为穿梭载体。它分为整合载体和自我复制载体两类。①整合载体：它带有一个酵母URA3标志基因和大肠杆菌的复制和报告基因。由于质粒DNA与酵母基因组DNA之间发生了同源重组,在转化的细胞中可以检测到质粒的整
cDNA
· cDNA Synthesis (Crawford Lab)mRNA can be converted into DNA (copy DNA, cDNA) by annealing oligo-dT to the 3' poly-A tail that occurs on
用低拷贝质粒作表达载体有什么好处
楼主的qq暂时加不上，我记下了，改天加。以下是个人意见。因为高拷贝质粒稳定性低，而且给宿主细胞的压力大。低拷贝数的质粒DNA在宿主细胞分裂前只能复制1～2次，而多拷贝数质粒可以在细胞分裂前复制成10～200拷贝。高拷贝数的质粒称为松弛型质粒(relaxed plasmid)，独立于细菌细胞而自主复制
293fectin™－Transfection
实验概要293fectin™ is a proprietary, cationic lipid-based formulation for transfection of DNA into eukaryotic cells. 293fectin™ is optimized for transfe
cDNA－Libraries
cDNA LibrariesIsolation of corresponding genetic informationInstead of synthesizing a desired gene, can we used the amino acid information to directly
Transient－transfection－into－293T－cells
PurposeTransient transfection into 293T cells is a convenient way to overexpress and obtain both cellular and extracellular (secreted or membrane) pro
BAC－EndSequencing
BAC End-Sequencing(Diana Bocskai)For every 4 mls of culture, dissolve the BAC DNA pellet in 40 µl of water. for example: Usually each BAC is grown in
Protein－Expression－and－Purification－Protocol
Step 1: Transform appropriate DNA plasmid into BL21(DE3) E. coli cells. These cells must be competent. (Protocol for how to make competent cells.)a) T
A－novel－method－of－growing－fungi－for－DNA－extraction
Preparation of fungi for DNA extraction typically involves growing cultures in liquid culture in Erlenmeyer flasks, Roux bottles or even microfuge tub
In－Vivo－Imaging－of－Far3
To determine the minimum dose of Katushka plasmid needed to give detectable fluorescent intensity, we decreased the amount of pTurboFP635 to 0.5 and 1
邓子新院士：实验室要营造利于创新的文化
5月29日下午，上海交大生命学院召开推进学院文化建设动员会。学院院长邓子新院士在讲话中，就如何开展实验室文化建设和学院教师分享了他多年的心得。邓院士认为，学校开展校园精神文化建设，生命学院应趁势而上，积极推进学院精神文化建设，无论把这个活动纳入全国的大框架，还是小到一个研究组，一个办公室，都是一个及
Twohybrid－analysis－of－genetic－regulatory－networks
1. Introduction and BackgroundThere is a great need for general methods to characterize the proteins that contemporary biology makes available. The li
Bacterial－Colony－PCR
Bacterial Colony PCRObjective:This protocol allows rapid detection of transformation success when primers are available to allow determination of corr
Agarose－Gel－Electrophoresis－of－DNA
1) Dissolve 1 g of agarose in 100 ml of 1X TAE or TBE buffer (gives a 1% gel). See note for making LMP agarose gel. 2) Cast the gel with the comb in p
细胞化学词汇质粒复制子
中文名称：质粒复制子英文名称：plasmid replicon定 义：在质粒DNA中能进行自主复制并维持正常拷贝数的一段最小的DNA序列。应用学科：生物化学与分子生物学（一级学科），核酸与基因（二级学科）
质粒复制子的定义
中文名称质粒复制子英文名称plasmid replicon定 义在质粒DNA中能进行自主复制并维持正常拷贝数的一段最小的DNA序列。应用学科生物化学与分子生物学（一级学科），核酸与基因（二级学科）
DNA转化实验指导2
1B. Cloning 1. A caveat on dephosphorylation: the most common reason for failure to obtain colonies is a result of adding too much BAP or CIP to
分子生物学实验小技术
当找到可能的重组质粒且条带清晰时可直接切下含DNA 的凝胶条带，用胶回收DNA 的方法回收质粒并进行酶切鉴定，如果DNA 量太少则需重提质粒。 用QIAprep Miniprep Spin 小量提质粒注意以下事项可提高得率A．用含抗生素的培养基培养细菌会有较高的得率。B．收菌时用tips吸干净残余的
酵母转化的几种方法
Modified Yeast Transformation Inoculate cells from an overnight culture into 50 ml YEPD and incubate at 30°C with shaking. Typically, add 0.1 to 0.2 m
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
CDNA文库
CDNA文库（主要内容如下）· Construction of cDNA Library· Construction of Genome DNA Library· Library Screening OthersConstruction of cD
人工转录因子的部件——人类锌指结构2
Table 2: Binding sites and identity of ZFPs used in VEGF activationWe then generated artificial transcription factors by fusing the three-finger domai
Performing－a－hunt－by－interaction－mating
AbstractWhen more than one bait will be used to screen a single library, significant time and resources can be saved by performing the interactor hunt
细菌人工染色体的概念
细菌人工染色体（Bacterial artificial chromosome，BAC）是指一种以F质粒（F-plasmid）为基础建构而成的细菌染色体克隆载体，常用来克隆150kb左右大小的DNA片段，最多可保存300kb个碱基对。
细菌人工染色体的特点
细菌人工染色体（Bacterial artificial chromosome，BAC）是指一种以F质粒（F-plasmid）为基础建构而成的细菌染色体克隆载体，常用来克隆150kb左右大小的DNA片段，最多可保存300kb个碱基对。