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RbCl Super Competent Cells

Xiyan Li Xiyan Li

Published: Jun 5, 2011 DOI: 10.21769/BioProtoc.76 Views: 24104

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Abstract

This method is used to inexpensively prepare home-made competent cells of E. coli. The transformation efficiency is at the high end of chemical-efficient competent cells, and close to library-efficient competent cells.

Keywords: Competent cells search E. coli search Chemical competent cells search Rubidium chloride search

Materials and Reagents

  1. 3 mM hexamine cobalt chloride
  2. Tryptone 
  3. Yeast extract
  4. NaCl
  5. DMSO
  6. KCl
  7. MgCl2
  8. MgSO4
  9. KOH
  10. CaCl2·2H2O
  11. RbCl
  12. SOB or 2x YT (see Recipes)
  13. MES (see Recipes)
  14. TFB (50 ml) enough for 80 tubes/400 transformations. DMSO (see Recipes)

Equipment

  1. Microcentrifuge

Procedure

Copyright: © 2011 The Authors; exclusive licensee Bio-protocol LLC.

Category

Microbiology > Microbial cell biology > Cell isolation and culture

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