The measures necessary to prevent PCR contamination in real-time PCR are broadly similar to those required for qualitative PCR. Due to the nature of the PCR process, contamination is a major potential problem. Contamination with PCR products may lead to false positive results. Unexpected positive PCR may result in the patient requiring re-sampling and cause unnecessary anxiety. There should be separate laboratories for specimen preparation, reagent preparation, PCR set-up and post-PCR processing. Although one of the major advantages of real-time PCR is that there is no need for post-PCR processing, many laboratories carry out qualitative PCR for the same target sequences. If it is not possible to have separate laboratories, careful consideration should be given to the design and utilization of the available space. Personal experience has shown that the majority of contamination problems occur during specimen processing where large numbers of specimens are being handled at one time.
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