Team:Calgary/21 June 2010


Monday June 21, 2010

Gel electrophoresis of Himika's construction of E1010 (RFP) and B0015 (Double terminators) as well as the restriction digest of the registry parts E1010 and B0015


Today I did another verification digest of the arabionose promoter (I0500). This time, I tried digesting with some none Biobrick restriction enzymes that have cut sites in the middle of the gene. Hopfeully, if this gives us bands of expected sizes, we can be sure that the arabinose promoter is what we think that it is. Today I also ran a gel of the verifictaion restriction digest that I perfomred on Friday. See the photo of the gel below.

Dev, Jeremy, and Chris

Today, Jeremy did gel electrophoresis of the restriction digest of his previous construct of B0015-R0040 using Biobrick restriction enzymes to test if the parts were correctly placed together. If the bands give show as expected, it is likely that the construct is what we expected it to be. The results of the gel electophoresis were inconsistent and as such, must be redone tomorrow. Chris did the construction of E0032-B0015 which is the end of the mRNA verification circuit. This was done by restriction digest, ligation, and transformation of the DNA into Top10 competent cells. Dev made overnight cultures of Registry parts J13002, J23032 and E0032 (kan plasmid transferred) so they could be Miniprepped tomorrow and ready for use.


Today, I did a restriction digest of E1010, B0015, and the construction of E1010-B0015. At first, I added 15 uL of the 10x Reaction Buffer but then redid it with the typical 3.5 uL. The gel results can be seen on the right. There were unexpected bands throughout as there was one band from E1010 at 5000 bp. The construction shows bands at approximately 200 bp instead of the expected size of 800 bp. This indicates that the construction was unsuccessful and must be redone.

Alex, Patrick, Raida

The restreaks of R0040-E0430 produced only one colony each. They did not glow yellow. The restreaks of the E0420 appear to be successful. They will be put into an overnight culture for tomorrow. We also reconstructed R0040 and E0430 because the one restreak did not appear to function well.