LargeScalePlasmidPreps:Qiagen/CesiumMethod

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 contaminating bacterial chromosomal DNA. If the chromosomal DNA is not sheared most of it will pellet along with the bacterial cell wall. However the relative purity of the plasmid DNA will drop if the plasmid is large or at a relatively low copy number in the prep. The co......阅读全文

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,

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 co

Large-Scale-Plasmid,-Cosmid,-BAC,-PAC,-and-Fosmid-DNA-Isolation

DNA Isolation by a Cleared Lysate Method Followed by Double Acetate Precipitation Version 3b - updated September 26, 1999The Most Recent Roe Lab Imple

质粒的大量制备

·         Plasmid Mini and Maxi Prep Methods (Gimila Lab) ·         Maxi-preps and all media, solutions (NWFSC)Isolation of cosmid, plasmid and P1 DNA

质粒的大量制备

·         Plasmid Mini and Maxi Prep Methods (Gimila Lab) ·         Maxi-preps and all media, solutions (NWFSC)Isolation of cosmid, plasmid and P1 DNA

Largescale-Immunocytology

This protocol describes our method for preparing cells for immunofluorescence, in which all incubations and washes are performed in microtiter dishes

Large-Scale-Tubulin-Preparation

Tubulin is purified from bovine/porcine brain by two cycles of polymerization/depolymerization followed by removal of copurifying proteins on a phosph

Large-Scale-Tubulin-Preparation——2

III. Pouring a 1L Phosphocellulose (PC) Column Resin: Whatman P11 Cellulose Phosphate -- fibrous cation exchanger (1 gram of PC swells to about 4 m

Small-scale-DNA-preps-for-Neurospora-crassa.

Molecular biology experiments often require preparation of small amounts of DNA from many samples. This abbreviated DNA isolation method yields an av

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

A-Method-for-MicroScale-Isolation-and-Purification-of-Gangliosides

A Method for Micro-Scale Isolation and Purification of Gangliosides Stephan Ladisch~Director, Center for Cancer and Transplant Biology, Children'

酵母转化

·         Yeast Transformation (Gietz Lab)LiAc/SS-DNA/PEG Transformation·         Yeast Transformation (Breeden Lab)LiAc method·         Large-Scale Y

One-step-miniprep-method-for-the-isolation-of-plasmid-DNA

plasmid miniprepAll ''miniprep'' methods reported so far for the isolation of plasmid DNA involve multiple pipetting, extraction, cent

质粒的小量制备

·         Standard (alkaline lysis) Mini-Prep (Goldberg Lab)Standard protocol for mini-prep and recipe for solution I, II and III.Alkaline Lysis Minip

质粒的小量制备

·         Standard (alkaline lysis) Mini-Prep (Goldberg Lab)Standard protocol for mini-prep and recipe for solution I, II and III.Alkaline Lysis Minip

General-Cloning-Protocols

Large Scale Preps: (See Large scale plsasmid prep protocol for more details) Cultures: Inoculate a 5 mL LB/Amp (50 - 100 µg/mL) culture in early a

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

BAC-DNA分离方法-Isolation-of-BAC-DNA-from-Largescale-Cultures

Isolation of BAC DNA from Large-scale CulturesJoseph SambrookPeter Maccallum Cancer Institute and The University of Melbourne, AustraliaDavid W. Russe

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

质粒的Midi制备

·         Plasmid Midi-preps (NWSFC)Diatomaceous Earth-based midi-prep. This procedure is the method of choice for isolating double stranded plasmid-b

细菌人工染色体

The Construction of Bacterial Artificial Chromosome (BAC) Libraries (complete manuscript) (Clemson University Genomics Institute)  Construction of BAC

DNA转化

DNA转化Chemical Transformation·         Transformation of Competent Cells (RbCl2 Method) (Goldberg Lab)Very nice protocol for E. Coli transformation inc

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

General-Laboratory-Procedures,-Equipment-Use,-and-Safety-Considerations

A. Storage . The following properties of reagents and conditions are important considerations in processing and storing DNA and RNA. Heavy metals pr

粘粒

Preparation of Cosmid DNAPreparation of Cosmid DNA from 50 ml Cultures (Donis Keller Lab)Cosmid vectors containing foreign DNA inserts are known to re

粘粒

Preparation of Cosmid DNAPreparation of Cosmid DNA from 50 ml Cultures (Donis Keller Lab)Cosmid vectors containing foreign DNA inserts are known to 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

CO2恒温摇床解决人胚肾-293-(HEK293)-细胞结团问题(一)

人胚肾 293 (HEK293)  细胞在重组蛋白表达中是最常见的宿主细胞。 这类细胞能够表达大量的膜蛋白,如 G 蛋白偶联受体  (GPCR) ,是无法在最常见的生物制药生产宿主,如:中国仓鼠卵巢 (CHO) 细胞中作表达。 HEK293 虽然是蛋白表达的极好宿主,然而 HEK293 细胞

使用CO2恒温摇床解决人胚肾-293-(HEK293)-细胞结团问题

人胚肾 293 (HEK293)  细胞在重组蛋白表达中是最常见的宿主细胞。 这类细胞能够表达大量的膜蛋白,如 G 蛋白偶联受体  (GPCR) ,是无法在最常见的生物制药生产宿主,如:中国仓鼠卵巢 (CHO) 细胞中作表达。 HEK293 虽然是蛋白表达的极好宿主,然而 HEK293 细胞

噬菌体的生长

Preparing Lawn Cells for M13 Cloning (Life Technologies)Lawn cells require the F' episome for M13 infection and may be prepared  Streaking Lambda