Bacterialglycerolstocks
To 2mls of mid-log culture or 1ml of freshly saturated culture add 1 ml(or an equal volume) of glycerol solution, mix gently, then freeze rapidly in liquid nitrogen or on dry ice. Store at -20°C and - 70°C.Glycerol solution: (Per 100 ml)65% glycerol (v/v) ............... 65 ml 100% glycerol0.1M Mg SO4 .......................... 10 ml 1M Mg SO40.025M Tris pH 8.0 ................ 2.5 ml 1M Tris22.5 ml ddH2Oautoclave to......阅读全文
Bacterial－glycerol－stocks
To 2mls of mid-log culture or 1ml of freshly saturated culture add 1 ml(or an equal volume) of glycerol solution, mix gently, then freeze rapidly in l
Streaking－Bacterial－Stocks
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
Bacterial－Media－Solutions－and－Stocks
3 agar (200 ml)Add 6 grams agar to 200 ml deionized water. Autoclave to sterilize.1.6 agar (200 ml)Add 3.2 grams agar to 200 ml deionized water.Autocl
细菌培养
Preparing Overnight Bacteria Culture (LaboratoryExperiments.com)This is a basic procedure for high school students and useful for those who are new to
Long－Term－Storage－of－Bacterial－Strains
Purpose:Bacterial strains may be stored indefinitely at low temperatures (- 20 degrees C and -80 degrees C) in 15 to 40 glycerol. It is lab policy to
Yeast－Media,－Solutions－and－Stocks
Yeast Media:Note: Synthetic complete medium can be prepared by adding media supplements (see below).Medium using 6.7 g yeast nitrogen base without ami
Bacterial－transformation
IntroductionTransformation is the process of introducing foreign DNA (e.g plasmids, BAC) into a bacterium. Bacterial cells into which foreign DNA can
DNA转化
DNA转化Chemical Transformation· Transformation of Competent Cells (RbCl2 Method) (Goldberg Lab)Very nice protocol for E. Coli transformation inc
XC－Assay－of－MoMLV－Virus－Stocks
MaterialsWildtype MoMLV virus aliquot. Stored at -80ºC.Medium: DMEM + 10% FBSNIH 3T3 TK- cellsXC cellsPolybrene 1000x stock = 4 mg/mL, sterile filte
Bacterial－Colony－PCR
Bacterial Colony PCRObjective:This protocol allows rapid detection of transformation success when primers are available to allow determination of corr
Bacterial－cell－culture
MaterialsGlass culture tubes with metal caps and labelsGrowth medium, from media room or customizedGlass pipette tubesParafilmEquipmentVortexerFireboy
Preparing－Overnight－Bacterial－Culture
Materials:Sterile LB medium (Luria-Bertani Medium) with or without antibiotic:water 500 mlbacto-tryptone 10 gbacto-yeast extracts 5 gsodium chloride 1
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
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
Glycerol－Agar－（甘油琼脂）的成分和适用范围
Glycerol Agar （甘油琼脂）Peptone （蛋白胨） 5g Beef extract （酵母膏） 3gGlycerol （甘油） 20g Top water （自来水） 1000mlAgar （琼脂） 15g pH 7.0-7.2
Bacterial－Expression－of－GSTfusion－Proteins
1. Grow cells in 5ml (or more) of LB-Amp overnight for a starter culture. 2. Grow larger culture (100x volume of starter culture) using the overnigh
Hydrolytic－Activity－of－Bacterial－Supernatants－for－Fungal－Suppression
As the fungal growth suppression by biocontrol agents (BCA) in solid media using dual plate assay has some issues regarding nutrient limitation. A pro
细菌培养基
Preparation of LB Plate (Dr. Chastain)prepare LB plate with or without antibioticsBacterial Culture Media Recipes (WUGSC) M9 Plate Supplement (Gottsch
Transformation－of－E.－coli－by－Electroporation
实验概要 Electrocompetent bacteria are prepared by growing cultures to mid-log phase, washing the bacteria extensively at low temperature, and
Methods－for－the－Measurement－of－a－Bacterial－Enzyme－Activity－in－Cell－Lysates
AbstractThe kinetic characteristics and regulation of aspartate carbamoyltransferase activity were studied in lysates and cell extracts of Helicobacte
TritonPrep－Method－for－bacterial－DNA－Purification
Triton-Prep Method for bacterial DNA PurificationGrow 5 ( large scale-15ml culture). Harvest in single eppendorf tube (or 15 ml disposable tube).Resus
细菌转化（bacterial－transformation）原理和操作
1．目的学会质粒DNA转化感受态受体菌的技术。2．原理质粒DNA粘附在细菌细胞表面，经过42°C短时间的热击处理，促进吸收DNA。然后在非选择培养基中培养一代，待质粒上所带的抗菌素基因表达，就可以在含抗菌素的培养基中生长。3．器材旋涡混合器，微量移液取样器，移液器吸头，1.5ml 微量离心管，双面微
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
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
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 untill the
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
Expression－Library－Screening－(Procaryotic)－Using－APFusion－Proteins
Outline:Bacteriophage lambda is a linear double stranded DNA, approximately 50 kB in size. The two sticky ends help the phage to recircularize after e
Preparation－of－Bacterial－Proteins－for－Analysis－by－2DPAGE
The following protocol has been developed for preparing soluble bacterial proteins in a form suitable for analysis by 2D-PAGE. The procedure was princ
Construction－and－Manipulation－of－LargeInsert－Bacterial－Clone－Libraries
Acknowledgements The organizer of the workshop acknowledges Dr. Murray Milford, Professor and Interim Head, and Dr. Mark Hussey, Professor and Interim
Production－of－Antibody－Fragments－in－Arabidopsis－Seeds
Plants offer a number of attractive benefits over conventional mammalian or bacterial cell culture systems for the production of valuable pharmace