User:Paul Rothenberg/Notebook/Pauls Notebook/2012/04/04

April 4, 2012

• Today, PCR was performed to sequence Myoglobin. The primers used were sw Mb M131Tf (reference number 84029666) and sw Mb M131Tr (reference number 84029667). In order for this to be completed, the primers were mixed with one mL of autoclaved water. Then, they were diluted with a calculated amount of sterilized water to bring the concentration to the correct amount for PCR. (See below).

• The protocol for PCR was downloaded from <microgen.ouhsc.edu/manuals_protocols/pcr_protocol.doc>, found on Google.

• Then, 2 microliters of DNA were added, along with 5 microliters of PCR buffer, 1 microliter of dNTP, 0.25 microliters of Taq Polymerase, 36.75 microliters of sterilized water were added, also including the two primers mentioned above.

• The mixture was then added to a temperature-controller, along with 25 microliters of wax to prevent evaporation. .