StreakingBacterialStocks
Principle:Bacterial cells are streaked onto a medium to obtain an independent isolate. This is done to reduce the likelihood of working with a culture which has become contaminated and/or has accumulated mutations.Time Required:Less than 5 minutes to streak out each strain; the colonies grow overnight.Procedure:The cells can be streaked from another plate a stab or from a frozen glycerol stock. Pick up a slightly vis......阅读全文
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
Lambda（噬菌体）DNA－Miniprep
David HarryInstitute of Forest GeneticsUSDA Forest ServicePacific Southwest Research StationAugust 26, 1993Background :There are many published method
美国实验室wetern方法
WESTERN BLOT PROTOCOLIn a Western blot, proteins that are separated on polyacrylamide gels on the basis of size are transferred to a membrane for dete
重组DNA的分离、克隆与测序实验手册3
G. Bacterial cell maintenanceFour strains of E. coli are used in these studies: JM101 for M13 infection and isolation (4), XL1BMRF' (Stratagene) f
Standard－Operating－Procedures－for－T1Phage－Testing－Assay
I. Introduction:This assay uses a lawn of phage-susceptible E. coli (DH10B) embedded in a layer of agarose. This top agarose lays on a bed of standard
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
384well－PCR实验方法
Beforehand.choose empirically the annealing temperature of primers;make sure you have enough PCR-plates, Q-covers and sealing tape;prepare the stocks:
Cosmid提取技术
Day OneStart overnight cultures of Cosmid in fresh L-Broth + 25 ug/ml kanamycin. 4 x 250 ml each, inoculate each with metal loop from frozen glycerol
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
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E. Elution of DNA fragments from agaroseDNA fragments are eluted from low-melting temperature agarose gels using an unpublished procedure first develo
Streptomyces:Protocols/Spore－Prep
Spore Prep - Inoculating & HarvestingDescription A spore prep is a method of preserving a sporulating strain of Streptomyces. The stock is stored in 2
酵母培养基和相关试剂
Yeast Hartwell's Complete (HC) Media (Gottschling Lab) Yeast Complete Media Yeast Media (PMCI) Yeast Media, Solutions and Stocks (Donis Keller
如何设计引物（二）
Nucleotides:Stocks of nucleotides for PCR (or other procedure) are NEARLY ALWAYS dNTPs (deoxynucleotides), and concentrations is almost always given i
DNA转化实验指导3
6. Simultaneous digestion of the pUC vector with both enzymes in the presence of 3 units of Shrimp Alkaline Phosphatase (Amersham BioSciences) in
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 ml saturated culture in the eve
Virus－Infection－of－Fibroblasts
Procedure: Day One 1. Plate cells in 60 mm dishes using 5 ml DMEM containing 10% Calf Serum. The protocol has worked well for NIH3T3 with 1 x 105 c
Virus－Infection－of－Fibroblasts
Procedure: Day One 1. Plate cells in 60 mm dishes using 5 ml DMEM containing 10% Calf Serum. The protocol has worked well for NIH3T3 with 1 x 105 c
Preparation－of－Plasmid－DNA－by－Alkaline－Lysis－with－SDS:－Minipreparation
实验概要Plasmid DNA is isolated from small-scale (1-2 ml) bacterial cultures by treatment with alkali and SDS.主要试剂Buffers and Solutions: Alkaline lysis
Agglutination－Assays
Agglutination Assays REFERENCE: Lanyi, B., and T. Bergan. Methods in Microbiology, Vol 10: 93-168. BACTERIAL AGGLUTINATION: Bacterial agglutinat
Preparation－of－Plasmid－DNA－by－Alkaline－Lysis－with－SDS:－Maxipreparation
实验概要Plasmid DNA is isolated from large-scale (500 ml) bacterial cultures by treatment with alkali and SDS.主要试剂Buffers and SolutionsAlkaline lysis solu
基本实验技术
I. Safety ProceduresA. ChemicalsA number of chemicals used in this laboratory are hazardous. All manufacturers of hazardous materials are required by
Fluorescence－Procedures－forthe－ActinandTubulin－Cytoskeleton－inFixed－Cells1
General StrategyWe typically work with tissue culture, primary mammalian cells, and cell extracts, but the protocols can be adapted to other systems,
亲和层析实验技术方法
INTRODUCTIONThis protocol describes a method for removing antibodies that react with bacterially encoded proteins by passing a crude preparation of im
Complement－Pathway
The complement pathway consists of a series of over thirty proteins in plasma that are part of the immune response. Activation of the complement syste
Cryopreservation－and－Recovery－of－Mature－Differentiated－Neural－Cells
实验概要Primary neuronal cultures are indispensable in the field of neurobiology and pharmacology. Many researchers favor freshly isolated neuronal cell
Preparation－of－phage－particles－from－phage－vectors
Pick up one phage vectors-containing colony with a sterile loop and put into 10 ml 2xTY + 10 µg/l tetracycline. Shake at 200 rpm and 37 °C unti
Transformation－Protocol－for－Arabidopsis
Transformation Protocol for Arabidopsis – AbbreviatedGerminate seed in pots↓ 4 weeksStreak bacteria onto YM/MinA↓ 2-3 days 28°CSpray/dip bacterial sus
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
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
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