Team:Penn State/Notebook

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Penn State iGEM 2012

Penn State Notebook

Monday, May 22, 2012
We prepared ampicillin and streptomycin plates in preparation for production of plasmids and competent cells.

Tuesday, May 23, 2012
Inoculation of LB broth with DH10B E. coli strain to prepare master mix/culture of cells in preparation for making chemically competent cells. These cells were incubated throughout the day and prepared for competency by the end of the day. Promotes were also planned out for the Multiple Promoters Project and ordered. These promoters were designed to combine some of the components of the plasmid.

Wednesday, May 24, 2012
Biobricks were hydrated and transformed into Chemically Competent cells through heat shock ( BBa_R0063, BBa_R1062, BBa_R0078, BBa_I1051, BBa_R0079, BBa_R0071 ). LOB broth was also prepared for the recovery from the transformation. These cells were then plated to produce isolated colonies.

Thursday, May 25, 2012
Additional Biobricks were hydrated and transformed into Chemically Competent cells through heat shock ( BBa_I712074, BBa_R0062, BBa_I746104, BBa_I719005, BBa_J23119, BBa_J23100, BBa_J23101, BBa_J23102, BBa_J23103, BBa_J23104, BBa_J23105, BBa_J23106, BBa_J23107, BBa_J23110 ). LOB broth was used to help the cells recover from their transformation. More agar plates were poured. The Transformed cells form yesterday yielded isolated colonies. A single colony from each plate was used to create a broth in preparation for mini-prep of the plasmids.

Friday, May 25, 2012
Miniprep was done to extract the first six plasmids we transformed ( BBa_R0063, BBa_R1062, BBa_R0078, BBa_I1051, BBa_R0079, BBa_R0071 ). Colonies that grew from the transformation of our last 14 plasmids ( BBa_I712074, BBa_R0062, BBa_I746104, BBa_I719005, BBa_J23119, BBa_J23100, BBa_J23101, BBa_J23102, BBa_J23103, BBa_J23104, BBa_J23105, BBa_J23106, BBa_J23107, BBa_J23110 ) were cultured for later plasmid extraction.

Saturday, May 26, 2012
Hannah and Kevin extracted the plasmids from the 14 colonies that were cultured yesterday. Two of the BioBricks that needed to be retransformed ( BBa_J23102, BBa_J23105) had isolated colonies on their plates and were cultured by Kevin for plasmid extraction the next day.

Sunday, May 27, 2012
Hannah and Kevin extracted the two cultures that Kevin prepared yesterday. These plasmids were put on ice for concentration analysis later with the other extracted plasmids.