The Optimization of RAPD Reaction System in Celery
In the present study, we set up separately five different concentration gradient to five main influence factors (dNTPs, Taq DNA polymerase, Mg2+, random primer and DNA template). Then based on the common RAPD reaction system, using the method of single factor progressive screening, we conducted the RAPD-PCR amplification of celery. Finally, we got the optimization of RAPD reaction system in celery. Conclusion of the optimized contents of 25 μL RAPD reaction system was as follows: Taq DNA polymerase 1.0 U, Mg2+ 3.0 mM,dNTPs 0.2 mM, primer 28 ng, DNA templet 70 ng, 10 × reaction Buffer 2.5 μL. The optimized PCR cycle program was as follows: 94℃ for 5 min, 94℃ for 1 min, 36℃ for 1 min, 722 for 1 min, 42 cycles and at final 72℃ for 10 min.
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NSTL
万方数据
维普
