Transformation Protocol Using Heat Shock
For ligation, leave tubes for 1 hour.) 9) Spread about 100 ul of the resulting culture on LB plates (with appropriate antibiotic added – usually Ampicillin or Kanamycin.) Grow overnight. 10) Pick colonies about 12-16 hours later. Title: Microsoft Word - …
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www.promega.comThe ligation conditions given in this protocol are based on the conditions used at Promega for quality control of lambda vectors with sticky ends. These ligation condi-tions have been developed using Promega Blue/White Cloning-Qualified T4 DNA Ligase. 3. The addition of polyethylene glycol (PEG) to ligation reactions can promote ligation of
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www.unl.educloning, including cleavage of DNA by many restriction enzymes, ligation of DNA, and bacterial transformation. * Because phosphates precipitate in ethanol, it is not possible to precipitate DNA and RNA from buffers that contain significant quantities of phosphate ions. * Phosphates sequester divalent cations such as Ca2+ and Mg2+.