Friday August 6, 2010

Jeremy

Today I ran my gradient PCR that I left overnight and the gel revealed that the pRFP did not get biobricked. Although this has been accomplished many times, for some reason this time it did not work. We had a meeting with out facilitators who suggested that I should try to increase the biobrick concentration because the bands are extremely faint for a PCR reaction. The thought was that the primers are not annealing as well as expected. The two suggestions were optimizing PCR or using a TOPO vector. Henry and Emily also suggested vacuum PCR purification to increase the concentration of DNA. Dr. Logan is also inquiring of GC richness in the DNA sample of pRFP. I also made the Notebook and Facilitators Page for the wiki and organized all the files for Patrick to put onto the wiki.

Dev

Colony PCR performed yesterday was unsuccessful. Plates with CpxR+I13504/I13507 construction were found in the fridge from Jeremy's construction in late June. Performed a colony PCR of 17 of these colonies. 8 of these colonies showed the correct size bands (850-900bp) on a 1% agarose gel.