Team:Calgary/16 June 2010


Wednesday June 16, 2010

Jeremy's and Dev's spread plates of the growth from the construction of J13002-E0040 and B0015-R0040

Dev, Jeremy, and Chris

Today, we observed the overnight cultures and the attempted constructions that were done yesterday. Dev spent the day registering for his courses. We Miniprepped the parts K239000, J61032, and K135000 which came from the Registry. The part J61032 was deemed unusable as the quality did not meet our standards as the plasmid concentration was observed. We also received periplasmic RFP from Dr. Schryvers' lab and the overnight cultures made from it were Miniprepped. As well, overnight cultures of many other parts from the Registry were made into glycerol stocks. Dev's construct of J13002-E0040 had tons of growth on the plate, but the colonies did not appear to fluoresce. Chris's plates of J13002-E0032 and pSB1K2-J23032 showed no growth once again. Tomorrow, another construction was planned with different vectors and inserts. As well, the Suffield R & D base was contacted and a possible tour was discussed with them.


Today I ran a gel of my PCR from yesterday. Alhtough there was some amplification, the bands were very faint, and it was hard to tell if it was successful or not. Colony 6 of the I0500-B0034 construct was sent down for sequencing with the BBK Sequencing primers. While waiting for sequencing, I've been looking into the sequence of the malE gene, which looks like it might be a vaery good candidate for our gene of interest in my circuit. The new I0500 part from the registry did not grow on either the plates or the broth cultures. Chris has contacted the registry and they are sending us yet another sample.


Both plates of growth of the construction of E1010-B0015 showed some growth. It is not certain that the parts are ligated together for sure, as there is no way to detect whether or not the vector closed on itself. Today, I did a restriction digest of the two parts and the construction to determine whether the part is present. Then, I did gel electrophoresis which showed one band in the lane with just E1010. Later, I will do a colony PCR and gel electrophoresis of the constructed part.

Alex, Patrick, Raida

The restreaks of K239000 and E0430 grew decently. We also miniprepped the E0430 part on pSB1K2, the E0420 part on pSB1A2, and K239000. We also did a gel electrophoresis of the colony PCR product from yesterday, in order to see whether the cpxP promoter was isolated properly. This gel failed, however, possibly due to some issues regarding the Taq polymerase used in the PCR reaction. We therefore retried the colony PCR using Platinum Taq.