Monday July 19, 2010

Emily

• Today I aliquoted our BBK-CP-F and BBK-CP-R primers. I then set up and ran a PCR with Taq Polymerase in order to test that our BBK construction primers are working. I also played around with the plate reader with our Arabinose induced cultures of I0500-B0034-E0040 from the weekend. Unfortunately, we didn't see any significant difference in GFP output between our cultures and a control of LB broth. We will start overnight cultures to retry the Arabinose induction procedure tonight. I also started overnight cultures of a J13002-E0040 construct in order to serve as a positive control in our GFP plate reading which we will be trying again tomorrow. Finally, I started a plasmid switch of our four constructs of I0500-B0034 in order to get them into a psb1AK3 vector and I also strated overnight cultures of B0034 in the psb1A2 plasmid, in order to start a plasmid switch of that construct tomorrow.

Patrick

Today, I set up and reupdated the team blog and Twitter. As well, I looked into papers regarding plasmid copy number and some basic JavaScript/CSS code to start constructing portions of the wiki.

Himika

Today I looked into plasmid copy number information which was assigned by Paul, who is the supervisor of our modeling component. I helped Emily set up the arabinose induced cultures but the readings of arabinose induction of I0500-B0034-E0040 versus LB broth(control) were not significantly different. Therefore, I believe that the RBS (b0034) did not get inserted as a result of GFP is not produced and readings are so similar. Due to this, I constructed I0500 with B0034.This is different from the previous constructions because this time I used I0500 as the insert so that it is easier to differentiate between the products that incorporated the insert versus products that did not. The products that incorporated the insert would be 1200 bp bigger than products that did not incorporate the insert. This is easier to screen.

Chris

Today, I redid the transformation of the CpxP promoter into both ampicillin-kanamycin and ampicillin-chloramphenicol plasmids with standard procedures. The digests were done on Thursday night but we forgot to plate them on Friday. Alex plated the tubes that were left shaking in the shaker over the weekend as well and we will see what kind of growth results tomorrow. As well, I got in contact with multiple companies that we were hoping for sponsorship such as PepsiCo, Sigma Aldrich, IDT, Finnzyme, and others. Tomorrow, if there is any growth with the CpxP promoters in plasmids, they will have overnight cultures made.

Raida

Today, I set up the PCR with Platinum Taq Polymerase to test whether our primers are working or not. This primer should anneal right outside the multiple cloning site and biobrick site, so our PCR product should have any biobrick part that is in the plasmid. I will run a gel of the PCR product tomorrow to see whether the primers have worked or not. I also plated the tubes that Alex was working on to give him a hand. For the Ethics portion, I read the notes that the team took from Suffield and the powerpoint that Suffield had prepared for us.

Jeremy

Today I plasmid switched I0500+B0034 from kan into chloramphenicol and gel extracted pRFP from its plasmid. I set up a PCR for the pRFP part. I also prepared a colony PCR of the DegP and CpxR tubes (there were 9 tubes) and set up part of the overnight for the same colonies that I PCR. Alex will be taking over the DegP and CpxR portions of the circuit. Cheers.