Abstract:

To establish RAPD reaction system of Cordyceps sinensis. Through changing the density of Mg2+, dNTP, primer, template DNA in RAPD reaction system, with the result of RAPD amplify, established the RAPD reaction system of Cordyceps sinensis. Then through changing the main thermal cycle parameter, optimize RAPD reaction system of Cordyceps sinensis. The results showed that an RAPD reaction system suitable for Cordyceps sinensis was established, that was: 20 μL amplification containing 1×PCR buffer, 1.5 μmol/μL Mg2+, 320 μmol/L dNTP, 24 ng premer、20 ng template DNA、1 U Taq DNA polymerase. The optimization result of amplifying the procedure was that: pre-Modified 5 min at 95℃, and then circulated 35 times (denatured 45 s at 94℃, renatured 1 min at 36℃, elongated 2 min at 72℃), and then elongated 2 min at 72℃ after the cycle. The RAPD reaction system can be used to RAPD analysis in Cordyceps sinensis.