From OpenWetWare
Jump to: navigation, search

Our Forward primer with the biobrick extentions arrived so a 100 micro molar stock was made and used to make a 10 micro molar dilution for use in PCR reactions. After reviewing the results of the gel electrophoresis from 10/18/11 it was noted that the band containg our PCR product were extremely bright indicating the concentration would be too high for continued reactions so a 1:100 dilution was made of our PCR product and then used to begin another PCR reaction using the forward and reverse primers with the biobrick extentions for future use in a biobrick compatable vector for cloning should they work. The samples were put in the thermocycler to complete the PCR reaction at a temperature of 58.8 degrees for annealing since the results from the previous PCR on gel electrophoresis indicated that temperature yeilded the highest results. Three samples were ran to icrease the volume of product obtained along with a negative control and two positive controls using a standard plasmid and the corresponding primers for that plasmid.

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