I've expressed Taq polymerase in E.coli. I have found a few reports for its purification also using sephadex columns. Next, I need to check its efficacy preferably by methods other than PCR. Can anybody suggest such functional tests?
Try to take one primer with annealing temperature to about 70o, and DNA for this primer (1-5 kb or moore), add 1mkCi P32 any of the 4 dNTPs to the reaction mixture, as for the sequencing, warm to about 94o (1-2 min) and place the reactions at 70-72o for 10, 30 and 60 minutes,. Then count the radioactivity.
Hi! Have you solved this problem? I have met the same problem , and I tried a method according to a paper which used Picogreen to maesure the dsDNA. Unfortunately, the results could not be stable, so I want to know much about the method measuring radioactivity, such as the amount of DNA ,the reaction system. How many is the least amount of radioactivity that can be detected. Waiting for your reply . Thank you !
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