840:153g:Projects/project24/2012/09/20

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Team Members: Sanju Timilsina and Parul Sirohi

Project title: Over expression of E. coli Acetyl- CoA carboxylase (ACC)sub-unit accC in E.coli to enhance fatty acid accumulation for Bio-fuel production”.

We prepared reagents ( Killing buffer, 10% SDS and 1X TE buffer) for DNA extraction on Tuesday Lab. We cultured our source organism E. coli 0157:H7 on 09/17 morning and incubated it for 28 hours. We performed DNA extraction from the cultured sample on 09/18 and stored our sample in ethanol and sodium acetate for 48 hours until next lab 08/20. We prepared three samples for agarose gel electrophorosis ( DNA sample 1, DNA sample 2 and DNA sample 1 digested with EcoR1). Under UV, we found that the band density of digested and undigested samples were similar, however the sample with EcoR1 digestion should give fragmented band and also the marker ( positive control) used didn't give good resolution so we decided to run the gel again in our next lab. We also decided to prepare the restriction enzyme digested sample for both DNA sample 1 and 2.

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