amplificationoccurs. The reaction mixture is heated to denature the isolated DNA and expose the primer targetsequences. As the mixture cools, the upstream and downstream primers anneal specifically to one of theDNA strands, Z05 extends the primer, and a second DNA strand is synthesized. This completes the firstcycle of PCR, yielding a double-stranded DNA copy of the target regions of the CT DNA and the CTInternal Control DNA. The reaction mixture is heated again to separate the resulting double-strandedDNA and expose the primer target sequences. As the mixture cools, the primers anneal to the targetDNA. Z05 DNA polymerase, in the presence of Mn2+ and excess dNTPs, extends the annealed primersalong the target templates to produce a double-stranded DNA molecule termed an amplicon. TheCOBAS® TaqMan® 48 Analyzer automatically repeats this process for a designated number of cycles,with each cycle intended to double the amount of amplicon DNA. The required number of cycles is preprogrammedinto the COBAS® TaqMan® 48 Analyzer. Amplification occurs only in the region of the CTcryptic plasmid and/or ompA chromosomal regions between the primers; the entire CT cryptic plasmidor chromosome is not amplified.
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