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Lambda Methods Media and Solutions

Updated Nov 29 1990

C. Helms


3% agar (200 ml)

1.6% agar (200 ml)

75 mM calcium chloride (100 ml):

DEAE-cellulose

DNaseI stock solution

0.5 M EDTA pH 8.0 (2 liters)

4 mM ferric chloride (100 ml)

2% gelatin (200 ml) 40% glucose (1 liter) Lambda diluent (100 ml) Lambda diluent + gelatin(100 ml) LB (1mM MgCl2 1 liter) 2X LB (1mM MgCl2 1 liter) LB plates LBM (150 ml) LB top agar (400 ml) LB+ Agar plates LB+top agar (1 liter) LMM medium (LB containing 1mM MgCl2 and 0.2% maltose) Large scale growth medium 1 M magnesium sulphate (1 liter) 1 M MgCl2 (1liter) Mussel glycogen (10 mg/ml stock) 3M potassium acetate 5 M potassium acetate (200 ml) 20% (w/v) PEG-8000 and 2.5M NaCl Proteinase K (20 mg/ml stock) RNase stock solution 10% SDS (100 ml) 2.5X SDS-EDTA dye mix (10 ml) 10 X SMO (1 liter) SM (1 liter) SM+ (1 liter) 3M sodium acetate (100 ml) TE (1 liter) X-Gal plates

References:

Arber W. Enquist L. Hohn B. Murray N. and K. Murray. (1983). Experimental methods for use with lambda.In: Lambda II (ed. R. Hendrix J.Roberts F. Stahl and R. Weisberg) pp. 433-466. Cold Spring Harbor Laboratory Cold Spring Harbor New York.

CRI Laboratory Manual: RFLPs Project (1989).

Helms C. Graham M.Y. Dutchik J.E. and M.V. Olson. (1985). "A new method for purifying lambda DNA from phage lysates". DNA 4: 39-49.

Sambrook J. Fritsch E.F. and T. Maniatis.(1989) Molecular Cloning A Laboratory Manual. Second edition. Cold Spring Harbor Laboratory Press.