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PCR of former temperatures
Due to the lack of success of our last PCR, we altered the annealing temperatures so that they would correspond with the temperatures used formerly (43, 48.5, and 52.1 C) so we could obtain a band at our target length. If we achieve the band at our target fragment length as we did with the first PCR, we plan to cut out this band and amplify the DNA in it to produce enough DNA to insert into a T-vector. If we are unsuccessful in our attempt, we will either sequence the band we amplify the most volume of or will pick up another project where a team has left off. Next time, we plan to run a 1% agarose gel electrophoresis to observe our success in terms of amplifying our target fragment length.