PreparingAntibioticsStockSolutionandAmpicillinAgarPlates
AMPICILLIN Beta-lactam-antibiotics are not very stable when dissolved. Slow but steady degradation happens even when frozen to -20°C. Therefore commercial beta-lactam-antibiotics are sold as dried powder and reconstituted just before use! If possible adjust stock solution pH to stability optimum and choose suitable storage temperature! Due to enhanced solubility Ampicillin-Na is used for cell culture instead of......阅读全文
Preparing Antibiotics Stock Solution and Ampicillin Agar Plates
AMPICILLIN Beta-lactam-antibiotics are not very stable when dissolved. Slow but steady degradation happens even when frozen to -20°C. Therefore comme
Preparation of Agar plates
Prepare media and add 1.5 agar before autoclaving it (15g per liter). After autoclavation, cool the media in a 55 degree waterbath. Do not allow th
Agar Plates for Selection of Clones in Bacteria
Cloning of PCR products Stocks: LB Agar: Luria Broth after Lennox: per Liter Tryptone 10 g Yeast Extract 5 g Sodium Chlori
PREPARATION OF 2% FORMALDEHYDE STOCK SOLUTION (2 METHODS)
METHOD 1:Formaldehyde preservative – 2% formaldehyde solution in protein-free phosphate-buffered saline (PBS).Prepare as follows:Add 2 g paraformaldeh
Expression Library Screening (Procaryotic) Using AP-Fusion Proteins
Outline: Bacteriophage lambda is a linear double stranded DNA, approximately 50 kB in size. The two sticky ends help the phage to recircularize afte
Preparation of Broth and Plates, etc.
Recipes: 1) LB Broth Make 16 gm of LB Broth Base (Gibco #M27800C) up to 800 ml in ddH2O. Swirl to dissolve, then add 110 µl of 10 N NaOH. Autoc
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
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
Maintenance of Probes in bacteria including Escherichia coli
Plasmid (pUC series) containing genomic DNA fragments are maintained in E. coli strain DH5aTM. The E. coli cultures are routinely cultured at 37 C on
Antibiotic concentrations for E. coli selection
Antibiotic ConcentrationsThe following list shows recommended antibiotic concentrations for LB media or agar plates.AntibioticConcentrationAmpicillin1
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
Preparing Lambda DNA
Preparing Lambda DNA1- Coliphage lambda DNA is a widely used vector for recombinant DNA. The middle third of its 48,000 bp contains no genes required
Pouring Plates
1. For rich media, weigh out appropriate ingredients and place into a flask. Add water until appropriate volume. Use a flask at least 2 times larger
Streptomyces:Protocols/Spore Prep
Spore Prep - Inoculating & Harvesting Description A spore prep is a method of preserving a sporulating strain of Streptomyces. The stock is stored
McKinney:TransformByElectroporation
Materials 50mL 7H9 mycobacterial medium + 3mL per transformation 102mL 10% glycerol (possibly a few mL more if you are doing many transformatio
Preparation of Bacterial Proteins for Analysis by 2D-PAGE
The following protocol has been developed for preparing soluble bacterial proteins in a form suitable for analysis by 2D-PAGE. The procedure was prin
Streptomyces:Protocols/Conjugation
Intergeneric Conjugation and Overlay Description Transfer of plasmid/cosmid DNA from a host strain, e.g. E.coli ET12567 [pUZ8002], to the recipient
Phage Titer
IntroductionLambda phage is a commonly used vector for transgenes. Its very high rate of infectivity makes it ideal for creating large numbers of clon
Expression Protocol in M9 Minimal Media via T7 Promoter
The following protocol has been used successfully to 15N or 13C/15N label our proteins using our pET1120/BL21(DE3) expression system: Preparing M9 min
Expression Protocol in M9 Minimal Media via T7 Promoter
Expression Protocol in M9 Minimal Media via T7 PromoterThe following protocol has been used successfully to 15N or 13C/15N label our proteins using ou
Inoue法制备大肠杆菌超级感受态细胞
实验步骤： 1、Inoculate from an overnight grown in LB.从培养过夜的LB平板上挑取单菌落 。2、Grow in 250 ml "SOB" at 18℃ until OD600 = 0.6.(0.3)接种于250ml SOB，18度培养至OD＝0.6。3、On
Easy Way to Clone Genes From a Phage Library
Easy Way to Clone The protocol is oriented towards a C. albicans genomic library I made in Lambda Zap II on 7/97. The overall sequence of ev
cDNA LIBRARY SCREENING
PREPARE SOLUTIONS1. 10mM MgSO4, 0.2% Maltose LB (100 mL):Mix 1.0 g of Bacto-Tryptone, 1.0 g of NaCl, 0.5 g of Yeast Extract, and 1.0 mL of 1M MgSO4. A
Hints and precautions for the care, feeding and breeding of Neurospora-2
9. Spontaneous germination. Some genotypes result in spontaneous ascospore germination (for example, the unpigmented ascospores of per-1 Type I). A s
Expression L19 using Pichia pastoris
Pichia pastoris is a methylotropic yeast used to express high amounts of protein. Secreted expression is achieved with a cleavalable faktor. To yield
Marcantonio Lab Protocol Manual——3
Sequencing Gel Preparing and Running a Sequencing Gel (6% Polyacrylamide/Urea) A. Preparation of Gel Solution 1) Weigh out 50 g of Urea into a
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
Dropout plates for yeast
Materials (Solutions are all available from the media room) 200ml bottle of 2x SD 200ml bottle of 4% agar -- make sure to sign it out 40% g
常用试剂和培养基
Sterile water bottles200 ml, 500 ml, or 950 ml water. Autoclave.0.05 M CaCl2 (per 200 ml)CaCl2·2 H2O 1.5 g Add 198 ml water.Autoclave.0.5 M CaCl2 (p
Preparing Overnight Bacterial Culture
Materials: Sterile LB medium (Luria-Bertani Medium) with or without antibiotic: water 500 ml bacto-tryptone 10 g bacto-yeast extracts 5