ransformation-Competent E. coli preparation

Rubidium chloride method Inoue "ultra-competent" method

Cosmid packaging protocol

DNA Ligation and Transformation Protocols

Medium and Buffers

Psi broth (per liter)compoundamountBacto yeast extract5 gBacto Tryptone20 g magnesium sulfate5 g pH 7.6 with potassium hydroxide

TfbI (per 200 ml) compoundamountfinal molarity/conc.potassium Acetate.588 g30 mM rubidium chloride2.42 g100 mMcalcium chloride0.294 g10 mMmanganese chloride2.0 g50 mMglycerol30 ml15% v/vpH 5.8 with dilute acetic acid

TfbII (per 100 ml)compoundamountfinal molarity/conc.MOPS0.21 g10 mMcalcium chloride1.1 g75 mMrubidium chloride0.121 g10 mMglycerol15 ml15% v/vpH 6.5 with dilute NaOH

Transformation "Ultra-Competent" E. coli (Inoue Method)

Inoue, H., H. Nojima, and H. Okayama. 1990. High efficiency transformation of Escherichia coli with plasmids.Gene 96:23-28. Citation Abstract