Abstract In this study,it established and optimized SSR detection system including
reaction components,amplified program,by using cabbage inbred lines 15R(‘Nanbao’)and
14S(‘Beiheidapingtou’).A stable SSR-PCR system was followed:10.00 μL reaction solution
contained 1×Buffer,50.00 μmol·L-1 dNTPs,0.40 U Taq enzymes,0.40 μmol·L-1 SSR primers
pairs,1.00 μL DNA(60.00 ng·μL-1).PCR amplifying program was divided into two steps,after
one denaturing step of 5 min at 94 ℃,DNA was denatured for 45 s at 94 ℃,the annealing
temperature was 60 ℃ and dropped 0.5 ℃ for each cycle,until a final annealing temperature
of 50 ℃ was reached.For the last 15 cycles of the amplification,an annealing temperature
of 55 ℃ was employed.When all cycles were finished,amplification ended for 10 min at 72 ℃
.The polyacrylamide gel electrophoresis was used in the detection of PCR amplification.
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