Haynes Lab:Notebook/Engineering PC-TFs/2014/03/20: Difference between revisions
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# Transfer all cells and DNA to respectively labeled SOC medium tube. Place back on ice. | # Transfer all cells and DNA to respectively labeled SOC medium tube. Place back on ice. | ||
# Tape both tubes horizontally to the shaking part (great science nomenclature) of the incubator for 30 minute recovery. | # Tape both tubes horizontally to the shaking part (great science nomenclature) of the incubator for 30 minute recovery. | ||
# Pellet cells by centrifuging at max rpm for three minutes. | |||
# Pipette out most of the SOC solution but leave a bit to re-suspend the cells. | |||
# Pipette volume onto labeled and warmed agar plates. Spread using sterile glass beads. | # Pipette volume onto labeled and warmed agar plates. Spread using sterile glass beads. | ||
# Incubate overnight at 37°C to get colonies | # Incubate overnight at 37°C to get colonies |
Revision as of 15:56, 20 March 2014
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Summary
Transform bacteria with the ligated plasmids 30 minutes
Note: The negative control will show you the number of “background” colonies so that you can determine whether your transformation worked, or is just the result of vector self-ligation or selection failure.
'Transforming bacteria with the ligated plasmids in BL21'
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