Week 1

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.

Week 2

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.

Monday, May 28, 2012
Memorial Day

Tuesday, May 29, 2012
The 20 plasmids that have been grow during the previous week and weekend were digested by Hannah, Victoria, and Kevin. The plasmids were run in a gel, however errors in the gel's preparation resulted in a poor run.

Wednesday, May 30, 2012
Plasmids for the Multidirectional Promoter Project were transformed and cultured by Hannah and Kevin. Chris, Kait, and Victoria re-transformed 10 plasmids and cultured those. A gel was run to confirm our plasmids and fragments but was inconclusive due to low concentration.

Thursday, May 31, 2012
Fragments for the Multidirectional Promoter Project were digested and run through PCR by Hannah and Kevin. Kait and Victoria re-transformed the remaining 10 plasmids and 1 from yesterday that did not yield colonies. Kait and Victoria ran a gel to confirm a plasmid they could use in the Codon Optimization Project.