Emily

This morning I helped make plates and I ran a gel of my PCR from yesterday. This PCR was using the M13 F/R primers that come with the TOPO Blunt cloning kit. As we had unfortunatley expected, there was no amplification, indicating once again that we do not have any inserts in our TOPO Blunt vectors. Raida and Henry are working on designing another PCR reaction to try to amplify malE, malE31, malESSdel and malE31SSdel out of our plasmid DNA and we will proceed from there. I was going to try the Arabinose Induction again, but I did a restriction digest of the plasmid DNA with EcoRI/PstI retsrcition enzymes of the I0500-I13504 and I0500-I13507 constructs, which revealed that the arabinose inducible promoter (I0500) was not able to be cloned in to the circuit. This means that the Arabinose Induction would be pointless, so I will abandon this attenpt until I have better verified constructs.

Chris miniprepped my colonies of malE and malE31 with the hypothetical XbaI and SpeI restriction sites. Because C2 was the only one that looked good from my colony PCR yesterday, I digested this with XbaI and SpeI again and ran this on a 1% agarose gel. It looked really good, as there was bands of the expected sizes: 1.4 KB for the insert, 3 KB for the vector and 4.2 KB for the insert and vector. I will proceed by doing one final verification digest of this colony with EcoRI/PstI and if that looks good, I will sned this colony down for sequencing as one final verification that it is finally biobricked!

This afternoon I also helped Chris and Raida PCR Purify some of their reactions from last week (CpxP and malESSdel). I also edited some sponsorship letters and worked on drafting up letters for the faculties of Kenesiology and Engineering.

Chris

Today, I used the vacuum purification method provided by Henry for purifying the PCR product of CpxP. I also started writing some more sponsor letters to the different faculties to attempt to get more funding for our team. I mini-prepped the colonies of malE and malE31 that Emily made last night. The plates of Dev's construction of K135000 into AK and AC plasmids showed little to no growth. The plasmids from the 2009 registry with the ccdB gene in them showed growth later on in the day and will be made into overnight cultures later.

Himika

Today I miniprepped the overnight cultures from last night. I also made overnight cultures for glycerol stocks of I0500-B0034 today. I also looked further into the literature for the modelling project.

Jeremy

My pRFP plasmid switch into pSB1AK3 and pSK1AC3 plates had growth. Instead of doing a colony PCR, I'm just doing overnights because there are very few colonies. I did 5 colonies from each type of plasmid switch and will miniprep and do a site directed mutagenesis of the internal PstI site later on. Today I primarily focused on the wiki, Paul has a meeting with Patrick and I. We talked about due dates which included having the main page organized and posted on the wiki when he comes back. I tweeked most of the main page and am starting to design the layouts for the other pages. We have finally decided on a template as a group!

Raida

Today, with the help of Henry, I set up and ran a PCR with the following templates (Plasmid containing MalE (P1HC1), Plasmid containing MalE31 (P31HC1), PCR products (linear MalE) and (linear MalE31)) with the following primers (Forward primer for MalE and MalE 31 with the MalE BBK-Reverse primer) and (Forward primer for MalE and MalE 31 with signal sequence deleted with the MalE BBK-Reverse primers)

I also continued reading some papers for the Ethics assignment.

Dev

Did not obtain colonies from yesterday's plasmid switch. Obtained a sample of CpxR in AK plasmid. Abandoned attempts to plasmid switch. Began the construction of CpxR in front of I13504 (GFP composite part), performed another construction of CpxR in front of I13507 (RFP composite part). Plated on NEW kanamycin plates.

Patrick

Today I discussed with Paul potential ideas for animation and modelling. I am now going to look into the coding of the wiki pages, with the help of Jeremy.