Transformation of E. coli with pKS/hly plasmid

Kama

• chemocompetent E. coli dH5α were incubated on ice for 15 minutes
• 15μl of ligation mixture was added (and 15μl of control ligation, without insert)
• bacteria were incubated with DNA on ice for 30 minutes
• heat shock was conducted (1 minute 42°C)
• bacteria were incubated on ice for 3 minutes
• After the heat shock 800μl of SOB medium was added
• Mixture with bacteria was incubated for 1 hour in 37°C
• 100μl of mixture was plated on medium containing ampicillin, X-Gal and IPTG
• The rest of mixture was rotated for 1 minute, pelet was

Ania

• Zupelnie nie chce mi sie nic robic jutro uzupelnie ten wpis a tymczasem fajny zel, wiki go docenilo i dalo mu numer

/6/66/Ania21.07.gif. A co u was?

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