Polymerase chain reaction (PCR) has been widely used to detect Mycobacterium tuberculosis for the rapid diagnosis of tuberculosis. In this study, FTC-2000 was used as a thermal cycler and the optimal reaction condition was established with it. I
used a
pair of primers based on IS6110 repeated sequence, which was reported to produce the 328-bps DNA products and known to be specefic to M. tuberculosis complex, and determined the optimal reaciton condition by the DNA band on the agarose gel
electrophoresis. The optimal conditions to improve PCR efficiency were 100μM of dNTPs each, 0.6μM of primers each, 2.5μg of BSA, 9.5 unit of Taq polymerase and 1.0 Mm MgCl2 in 10μl reaction mixture. And the denaturation time could be reduced
to
1
sec at 94℃. The 40 cycles reaction was needed for the detectable 328-bps DNA band, but the reaction over 60 cycles was needed for clear DNA band. Under these conditions, the PCR using FTC-2000 may help the rapid detection of the M. tuberculosis.
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