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Team:UTK-Knoxville/17 June 2013
From 2013.igem.org
June 17th, 2013
While Eric is preparing chemi competent cells for transformation Brandon ligated Aer-EnvZ +OmpR + backbone again using the same digested pieces Eric used.
3:1 - 1 uL BB, 1 uL Ligase, 1 uL Buffer, 1.5 uL OmpR, 1.5 uL Aer/EnvZ, 4 uL sterile DI water.
5:1 - 1 uL BB, 1 uL Ligase, 1 uL Buffer, 3 uL OmpR, 3 uL Aer/EnvZ, 1 uL sterile DI water.
Ligated for 7 hours at room temperature.
Patrick transformed the ligations into TOP10 cells. Used LB as a recovery medium. Then added 250 uL of 500 g/L glucose at 30 minutes into recovery. Plated cells both concentrated and not. A small pellet was formed after centrifuging, so some cells may grow.
