Convective polymerase chain reaction in standard microtubes

Polymerase chain reaction (PCR) is the most widely used method for nucleic acids amplification. To date, a huge number of versatile PCR techniques have been developed. One of the relevant goals is to shorten PCR duration, which can be achieved in several ways. Here, we report on the results regarding nucleic acids amplification by convective PCR (cPCR) in standard 0.2 ml polypropylene microtubes. The following conditions were found to be optimal for such amplification: 1) 70 mu l reaction volume, 2) the supply of external temperature 145 degrees C for the denaturation zone and 0 degrees C for the annealing zone, 3) similar to 30 degrees inclination of the microtube main axis, 4) the use of nearby primers, and 5) duration of the reaction 15-20 min. At these conditions, the amplification products are accumulated in an amount sufficient to be registered by gel electrophoresis, and high sensitivity of the reaction comparable to that of conventional PCR is achieved. cPCR provided the reliable detection of SARS-CoV-2 coronavirus RNA isolated from nasopharyngeal swabs of COVID-19 patients.

Automated summary

Polymerase chain reaction (PCR) is widely used for nucleic acids amplification. This study reports the use of convective PCR (cPCR) in standard microtubes for efficient amplification of nucleic acids. By optimizing reaction conditions, high sensitivity detection and accumulation of amplification products were achieved, enabling reliable detection of SARS-CoV-2 coronavirus RNA from COVID-19 patients' nasopharyngeal swabs.