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. Always make sure your gasket is correctly seated before you pour the agarose!

Wednesday, May 30, 2012
Plasmids for the Multidirectional Promoters 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.
The originally transformed promoters were disposed of. Some of the cultures became a purple color. After centrifuging it was shown that the bacteria themselves were purple. This may be from contamination, so to be on the safe side, these plasmids were re-transformed. This may be an artifact from the plasmids themselves, but there is no consistent pattern. Very strange indeed! Always remember to use good sterile techniques when working with media and cultures!

Friday, June 1, 2012
Today Hannah digested components for the Multiple Start Codon Project. Later she ran a gel and extracted the components for later use. Kait and Victoria prepared a plasmid for the Codon Optimization Project. Kevin and Chris began work on a plasmid backbone and fluorescent component of the Multidirectional Promoters Project.
The team also received a new gel illuminator to see their bands without the use of UV light. They were very excited.

Week 3

Monday, June 4, 2012
Hannah hydrated BBa_I13521 and used it to gauge the competency of the chemically competent cells we prepared earlier. Victoria and Kait ran a gel to confirm some of the components they need for the Codon Optimization Project. Kevin and Chris worked on some cleaning and housekeeping.

Tuesday, June 5, 2012
Victoria and Kait completed a transformation for the Codon Optimization Project. Kevin mini-prepped two plasmids for the Multidirectional Promoters Project. Chris and Kevin then froze some cultures containing these plasmids for later use. Hannah completed a CBAR and transformation for the Multiple Start Codon Project. She also tested the competency rate of the chemically competent cells the team prepared earlier.