| 摘要 |
<p>A method for amplifying a target nucleic acid sequence by convective polymerase chain reaction (PCR), comprising the steps of: 1. providing a 5 tube-like container; 2. placing a PCR sample in the tube-like container, where the sample contains a template DNA, DNA polymerases, deoxyadenosine triphosphates (dATPs), deoxycytidine triphosphates(dCTPs), deoxyguanosine triphosphates (dGTPs), deoxythymidine triphosphates (dTTPs), and at least two oligonucleotide primers having a sequence complementary to the 3' end of the target nucleic acid sequence where the primers are designed to have a melting temperature (Tm) between about 40°C to about 90°C; 3. embedding a bottom part of the container in a heat source, and then heating the PCR sample to allow the primers melt and maintaining a steady temperature in the surface of the PCR sample (Ts) of at least 2 °C less than the melting temperature of the primers (Tm) and controlling the following parameters of the PCR comprising: total volume of the PCR sample (V), viscosity in Ns/m2 of the PCR sample (ì), inner diameter in mm of the container (d), and the surface temperature in °C of the PCR sample in the container (Ts); so that a temperature gradient descending from bottom to top is resulted in the sample, which induces a convection and makes the following PCR events (i) denaturation (ii) annealing and (iii) polymerization yield a copied DNA template</p> |
