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Transformation of Bacteria

190 bytes added, 15:05, 5 May 2009
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*Plasmid amplification use subcloning efficiency DH5a
*Cloning use OneShot TOP10 (Pink)
*Mutagenesis use XL1 Blue Supercompetent (Blue), comes in 50uL aliquots
*SOC Buffer
*DNA of interest (1 uL for plasmid, 5 uL for cloning/mutagenesis)
#Heat shock at 42C for 45s
#Place back on ice
#Add 450 uL of SOC Bufferor LB on shelf OR add 1000uL and adjust the level.
#Incubate at 37C for 1h with occasional mixing
#Plate 30-50 uL for amplification, or all for cloning/mutagenesis*When plating be sure to label plates and use extreme caution when sterilizing the spreader with EtOH and fire.
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