Wednesday August 4, 2010'

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.

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.

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.