Himika's gel electrophoresis of the restriction digest of parts listed below

Jeremy's construction of R0040+ 13507. Red and Normal colored cells present.

Jeremy's Bbk RFP (attempt 1)

Friday July 9, 2010

Himika

Today I did a restriction digest of parts with EcoRI/SpeI in React II Buffer that I constructed and the plasmid switch I did on July 6th. The parts that I digested were:

• 3 colonies of E1010 in AC plasmid
• 2 colonies of E1010-B0015 construction done on July 6th
• I also ran 2 colonies of I13507
• 1 colony of E1010 in its original Kanamycin plasmid

Refer to the image for the results of the gel electrophoresis done of the restriction digests

I also designed primer along with Dave for sewing PCR of the ppRFP circuit.

Chris

Today, I contacted the Qiagen representative about possible sponsorship and am awaiting a response. As well, this morning, I did plasmid preparations of several different overnight cultures of I0500. 7 overnight cultures were prepped using the Sigma Aldrich GenElute Plasmid Preparation kit (the protocol can be found in our Lab Protocols section).

Jeremy

Today I worked on biobricking pRFP because the primers came in. Henry helped design suitable conditions for the PCR. There was problems with larger than normal bands, potentially due to annealing on the plasmid. The R0040 + I13507 parts had colonies that are red, further verification via sequencing for I13507 will be necessary to confirm it is consistent.

Dev

Results: Growth was obtained. Colonies showed fluorescence.

Alex

Today our new reverse primer for cpxp arrived. I diluted the primer down to ~5 micro molar, and then also diluted the forward primer, getting close with about 7 micro molar. Then we prepared a 5X master mix with our primers, and ran 5 PCR reactions. Today we also presented our project to our supervisors.

Team

We did a presentation of our overall project to our supervisors Dr. Cairine Logan and Dr. Anthony Schryvers as well as several graduate students and our lab technician Deirdre Lobb. This presentation was put together as a practice for the iGEM competition as well as to inform everyone on the status and goals of our project. Changes to both the project as well as the presentation were suggested by our supervisors and modifications will be made. One of the main points which were made was the need for clarity in all slides. What we thought was easily understandable in figures turned out not to be so to others. Visual depictions of circuits and colour schemes will need to be modified in the future.