Gel electrophoresis of Jeremy's construction of B0015-R0040 and Himika's construct of E1010-B0015

Friday June 18, 2010

Emily

Today I did a restriction digest of the arabinose promter (I0500) using all combinations of restriction enzymes. The sequencing of this part came back very strange, so I want to verify the presence of all of the Biobrick cut sites within the part. Today I also started a colony PCR using the BBK Construction primers of my I0500-B0034 construct that I constructed yesterday.

Dev, Jeremy, and Chris

Today, Jeremy did gel electrophoresis of different colonies from his construction of B0015-R0040 and Himika's construct of E1010-B0015 which is shown in the photo on the right. The bands observed were much clearer than those of the gel done yesterday. We had a group meeting in the afternoon to discuss with Dr. Logan and Deirdre about the progress during the week. Dr. Logan pointed out a cheaper method to verify whether the DNA was present by searching for restriction sites within the DNA, digesting with them, and checking the size of the parts with a gel electrophoresis. Chris's plates from yesterday of J13002-E0032 and pSB1K2-J23032 showed no growth once again. They will be attempted again next week with the vector and insert reversed. Dev had growth on his construction of J13002-E0040 but the growth did not exhibit green fluorescence.

Himika

Today, I did Miniprep Plasmid Preparations of the overnight cultures made yesterday of the construct of E1010-B0015 using the Sigma Aldrich Plasmid Preparation kit. The plasmid concentrations were decent and the purifications as measured with a spectrophotometer. They will be restriction digested and run on a gel to determine their viability of use on Monday.

Alex, Patrick, Raida

The E0420 plasmid switch appeared successful. There are a lot of colonies, but we will check for fluorescence on Monday. We restreaked the spread plates of E0420 and the R0040-E0430 construct.