User:Madeline Hartman/Notebook/Dr. Hartings Lab/2012/11/27: Difference between revisions
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==Entry title== | ==Entry title== | ||
Cell Transformation was performed | |||
# BL21(DE3) cells thawed gently on ice | |||
# The competent cells were mixed. 100 μL cells were added to three separate 14-ml BD Falcon polypropylene round bottom tubes. | |||
# The three tubes were incubated on ice for 10 minutes, swirling gently ever 2 minutes | |||
# 5 μL, 10 μL, and 15 μL of ligated DNA were added to one of the three tubes, and swirled gently | |||
# The three tubes were incubated on ice for 30 minutes | |||
# Each of the three tubes was heat-pulsed for 45 seconds in a 42°C water bath | |||
# The three tubes were incubated on ice for 2 minutes | |||
# 0.9 ml of LB medium was added to each of the three tubes and the tubes were incubated at 37 °C for 1 hour while shaking at 225 rpm. | |||
# 200 μL of cells were plated and spread with a sterile spreader | |||
# The cells were incubated overnight at 37°C | |||
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Revision as of 12:59, 4 December 2012
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Entry titleCell Transformation was performed
|