915
edits
Changes
copied over protocol
==Materials==
*Competent Cells
*Plasmid amplification use subcloning efficiency DH5a
*Cloning use OneShot TOP10 (Pink)
*Mutagenesis use XL1 Blue Supercompetent (Blue)
*SOC Buffer
*DNA of interest (1 uL for plasmid, 5 uL for cloning/mutagenesis)
*Plates (Amp or Kan; in cold room)
==Protocol==
#Thaw cells on ice and label
#Add DNA to cells and mix by tapping
#Incubate on ice 30-45min
#Heat shock at 42C for 45s
#Place back on ice
#Add 450 uL of SOC Buffer
#Incubate at 37C for 1h with occasional mixing
#Plate 50 uL for amplification, or all for cloning/mutagenesis
*Competent Cells
*Plasmid amplification use subcloning efficiency DH5a
*Cloning use OneShot TOP10 (Pink)
*Mutagenesis use XL1 Blue Supercompetent (Blue)
*SOC Buffer
*DNA of interest (1 uL for plasmid, 5 uL for cloning/mutagenesis)
*Plates (Amp or Kan; in cold room)
==Protocol==
#Thaw cells on ice and label
#Add DNA to cells and mix by tapping
#Incubate on ice 30-45min
#Heat shock at 42C for 45s
#Place back on ice
#Add 450 uL of SOC Buffer
#Incubate at 37C for 1h with occasional mixing
#Plate 50 uL for amplification, or all for cloning/mutagenesis
