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Transformation Protocol: RbCl-method Media and Solutions
 

Summary: Transformation Protocol: RbCl-method
Media and Solutions
YETM, 500 ml: 2.5 g Yeast Extract
10 g Tryptone
5 g MgSO47H2O
Adust pH to 7.5 with KOH. For plates: add 7.5 g of Agar
TFB1, 200 ml: 0.59 g KOAc (30 mM)
2.42 g RbCl (100 mM)
0.29 g CaCl22H2O (10 mM)
1.98 g MnCl24H2O (50 mM)
30 g Glycerol (15% wt/vol)
Adjust to pH 5.8 with 0.2 M acetic acid (do not adjust pH with KOH). Add dH2O to 200 ml. Filter
sterilize. Store refrigerated at 4C.
TFB2, 200 ml: 0.42 g MOPS (10 mM)
2.21 g CaCl2-2H2O (75 mM)
0.24 g RbCl (10 mM)
30 g Glycerol (15% wt/vol)
Adjust to pH 6.5 with KOH. Add dH2O to 200 ml. Filter sterilize. Store refrigerated at 4C.
Preparation of Competent Cells
1. Streak cells from frozen stock onto YETM plate. Incubate overnight at 37C.

  

Source: Aris, John P. - Department of Anatomy and Cell Biology, University of Florida

 

Collections: Biology and Medicine