High Molecular Weight Yeast Liquid DNA Preparation

Solutions:

 

• SCE

  SCE		Final concentration
  2M Sorbitol	50 ml	1.0 M
  1M Sodium citrate	10 ml	0.1 M
  0.25M EDTA, pH 7.0	24 ml	60 mM
  sterile ddH2O	16 ml
  	-------
  	100 ml

  Filter sterilize and store at room temperature.

   

   

• Large Scale Prep Lysis Buffer:

   

  0.5 M Tris-HCl , pH 9.0

  3% Sarkosyl

  0.2 M EDTA

   

References:

Carle, G.F., and M.V.Olson. (1984) "Separation of chromosomal DNA molecules from yeast by orthogonal-field-alteration gel electrophoresis." Nucleic Acids Res. 12:5647-5664.

Burke, D. T., Carle, G. F., and M.V.Olson."Cloning of large segments of exogenous DNA into yeast by means of artificial chromosome vectors." (1987) Science 236:806-812.