Team:Calgary/5 September 2010


Sunday September 5, 2010

Chris's gel of the restriction digest of all old CpxP and ibpAB stuff. Lane 10 and Lane 4 show possible cuts but all other lanes show nothing. Lane 10 is ibpAB and Lane 4 is CpxP3 but those two tubes have no more DNA and must be redone.

Who doesn't love weekends?


Today, I ran a gel of the restriction digest of all the CpxP and ibpAB stuff from yesterday which was done with NEB enzymes. The gel results can be seen on the right. ibpAB1 (Lane 10) showed digestion at about the right size, but there was no more in the tube to construct with. As a result, overnight cultures were made of it. Another digest was set up of all the CpxP and ibpAB stuff that was ligated into Biobrick plasmids and a positive control of a Lux circuit from last year was also digested to affirm the enzyme activity. Finally, I helped Emily set up her multi-salt concentration PCR of MalE once again.


Today I ran a gel of my colony PCR's from yesterday of malE and malESSdel. Unfortunately, I didn't get any amplification. So I proceeded to set up three PCR reactions of malE and malE31, varying the concentration of mgcl2 from 1.5 to 2.5 as well as running a temperature gradient from 51 to 59 degrees celcius.


DO NOT WORRY BLOG READERS, I AM STILL ALIVE! Anyway, today I worked on the construction of CpxR + I13504 and CpxR +I13507. The construction was ligated into the AK plasmid, transformed and plated on AK plates.