840:153g:Projects/project17/2011/10/13

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Today a new restriction enzyme digest was set up since the dye was forgotten in the gel during the previos lab period, as well as, incorrect running of the electrophoresis of the gel. The PCR reaction that was set up and ran through the thermocycler on 10/11/11 was run on a new gel along with the repeated RE digest and a picture was taken for both of the results. The PCR reaction showed that our primers cut out the correct gene at approximately 1185bp and worked at all three temperatures of 48, 53, & 58 degrees. The reaction at 58 showed the largest amount of dna but also had a faint extra band. The bands were cut out of the gel, divided in half, and stored in the -80 degree freezer in 2 separate centrifuge tubes to be used after ordering the forward primer with biobrick extentions in addition to the reverse primer already stored with the biobrick extentions inorder to proceed with a biobrick compatable vector. The RE digest also showed that all four restriction enzymes were functional with all three of the promoter parts ordered.

Reference article: Evolution of steroid-5-alpha-reductases in comparison of their function with 5ß-reductase DOI: 10.1016/j.ygcen.2009.08.004

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