The factors influencing RAPD Analysis, including the concentration of DNA template, Mg2+, dNTP, primers, Taq polymerase, Thermal cycles and annealing temperature in tobacco were studied. An optimal PCR system for RAPD in tobacco was found: in 25μl reaction solution, contained 40ng DNA template, 0.4μM random primers, 2.5mM Mg2+, 0.2mM dNTP, 1U Taq polymerase. The amplification program was devised: 94℃ for 5min;denaturing at 94℃ for 1min; primer annealing at 38℃ for 1min, extension at 72℃ for 1.5min, 39 cycles; at last extension at 72℃ for 5min.
Yang Youcai, Zhou Qingmin, Yin Hanqi
. Establishment and Optimization of RAPD Reaction System in Tobacco[J]. Chinese Agricultural Science Bulletin, 2005
, 21(5)
: 97
-97
.
DOI: 10.11924/j.issn.1000-6850.050597
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