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
细菌人工染色体
The Construction of Bacterial Artificial Chromosome (BAC) Libraries (complete manuscript) (Clemson University Genomics Institute) Construction of BAC
重组质粒（dna－recombinant－plasmid）的连接、转化及筛选1
第一节 概 述质粒具有稳定可靠和操作简便的优点。如果要克隆 较小的DNA 片段(＜10kb)且结构简单,质粒要比其它任何载体都要好。在质粒载体上进行克隆 ,从原理上说是很简单的，先用限制性内切酶切割质粒DNA 和目的DNA 片段, 然后体外使两者相连接, 再用所得到重组质粒转化细菌,即可完成
重组质粒（dna－recombinant－plasmid）的连接、转化及筛选2
第二节 材料、设备及试剂一、 材料外源DNA 片段: 自行制备的带限制性末端的DNA 溶液,浓度已知; 载体DNA : pBS质粒(Ampr ,lacZ),自行提取纯化,浓度已知; 宿主菌: E. coli DH5α,或JM系列等具有α-互补能力的菌株。二、 设备恒温摇床，台式高速离心机，恒温水浴锅
Infusion－biobrick－assembly
OverviewThis is a method to assemble two BioBricks using the Clontech In-Fusion PCR Cloning Kit and maintains BioBrick standard formats. There are cur
DNA克隆
DNA克隆（主要内容如下）· General Procedure· PCR Cloning· Subcloning· ET Cloning· Vector Preparation· Ligation Re
重组DNA的分离、克隆与测序实验手册8
B. Midiprep double-stranded DNA isolationA midi-prep double-stranded DNA isolation has been developed to generate a sufficient amount of template DNA
关于黏性质粒载体的简介
由于真核基因的结构与功能研究的需要，人们发展出比λ噬菌体载体具有更大克隆能力的新型的载体——柯斯质粒载体(cosmid vectors，cosmid是COS site—carrying plasmid的缩写)，也称为黏陛质粒或黏粒。这是一类人工构建的含有抗性基因、单一克隆位点以及λDNACOS位
关于黏性质粒载体的基本介绍
由于真核基因的结构与功能研究的需要，人们发展出比λ噬菌体载体具有更大克隆能力的新型的载体——柯斯质粒载体(cosmid vectors，cosmid是COS site—carrying plasmid的缩写)，也称为黏陛质粒或黏粒。是基因工程中一个重要的基因表达载体。
Streptomyces:Protocols/MiniMaxi－Prep
Small Scale Plasmid Isolation (Mini / Maxi Prep)Description A mini prep / maxi prep is used to isolate plasmid or cosmid DNA from bacteria, normally E
Streptomyces:Protocols/Transformation－by－Electroporation
Description Transform E.coli cells with plasmid/cosmid DNA using the method of electrophoration (inserting plasmids into E.coli).Approx. Duration:Prep
Transfecting－Plasmid－DNA－into－NIH3T3－Cells－Using－Lipofectamine™－LTX－Reagent
实验概要Lipofectamine™ LTX Reagent is a proprietary, animal-origin free formulation for the transfection of DNA into eukaryotic cells with low cytotoxic
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
Miniprep/Kitfree－highthroughput－protocol
BackgroundThis protocol is adapted from "Molecular Cloning: A Laboratory Manual", Second Edition, Sambrook, Fritsch, and Maniatis. It is a quick, inex
M13噬菌体
· M13 Phage (Michael Blaber)Very useful background information about M13: its infection, replication, packing, cloning. If you are new to phag
DNA纯化手册1
Purification of plasmid DNA (miniprep) with high yields using diatomaceous earthKyung-Soo Kim and Charles K. PallaghySchool of Botany, La Trobe Univer
柯斯质粒载体的特点
柯斯质粒是一类人工构建的含有lDNA的cos序列和质粒复制子的特殊类型的质粒载体，cosmid是cos site carrying plasmid的缩写。柯斯质粒的大小为4-6kb，由3部分组成：A.多克隆位点区B. 含有cos位点的lDNA区C. 复制起始位点和抗性标记区
重组DNA的分离、克隆与测序实验手册4
H. Fragment purification on Sephacryl S-500 spin columnsDNA fragments larger than a few hundred base pairs can be separated from smaller fragments by
Sequencing－off－Cosmid,－BAC,－PAC,－－with－ABI－Big－Dye－Terminators
Big Dye Protocols and Notes - Cosmid, BAC, BAC, Fosmid TemplatesHi all,Over the past two months, we have been testing various reaction conditions for
Fastfilter－Plasmid－Midi－Kit－Spin－Protocol
实验概要The E.Z.N.A.TM Fastfilter Plasmid Midi Kit combines the power of HiBind® technology with the time-tested consistency of alkaline-SDS lysis of
Large－Scale－Plasmid－Preps:－PEG－method
1. Grow 250 500 mL of bacteria overnight in LB with 50 µg/mL of ampicillin.2. Transfer culture to Nalgene bottles. Spin in SGA rotor at 6000 rpm, at
DNA纯化手册2
Key points to observe: a. Use a endA1- E. coli strain for plasmid propagation and isolation whenever possible. The instability of plasmids isolated fr
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
Large－Scale－Plasmid－Preps:－Qiagen/Cesium－Method
Most plasmids can be adequately prepped by kits containing DNA binding columns. These columns do not do a great job of separating plasmid DNA from con
Fastfilter－Plasmid－Midi－Kit－Vacuum/Spin－Protocol
实验概要The E.Z.N.A.TM Fastfilter Plasmid Midi Kit combines the power of HiBind® technology with the time-tested consistency of alkaline-SDS lysis of
Bacterial－cell－culture
MaterialsGlass culture tubes with metal caps and labelsGrowth medium, from media room or customizedGlass pipette tubesParafilmEquipmentVortexerFireboy
Ligation－Optimization
The following protocol can be used to optimize ligation conditions for difficult to clone (e.g. very large) fragments. The principle is to independent
E.Z.N.A.™－Fastfilter－Plasmid－Mega－Protocol
实验概要The E.Z.N.A.™ family of products is an innovative system that radically simplifies extraction and purification of nucleic acids from a variety
Lambda噬菌体
· Lambda DNA Preparation (Stanford DNA Sequence & Technology Center)Detailed protocol for lambda DNA preparation with recipes· Isolati
Basic－procedures－for－bacteria－culture2
E. Elution of DNA fragments from agaroseDNA fragments are eluted from low-melting temperature agarose gels using an unpublished procedure first develo
E.Z.N.A.®－Plasmid－Maxi－Kit－Spin－Protocol
实验概要This Protocol is designed to isolate 500-1200 ug of high Copy-Number plasmids or 50-400 ug of low Copy-Number Plasmids from 200 ml overnight c