SCoT-PCR optimization in Halocnemum strobilaceum L.

In this research, the PCR optimal conditions were studied for ۱۲ primers of SCoT in Halocnemum strobilaceum L. Five parameters including, the temperatures and times for annealing and extension, PCR cycle numbers, the concentration of template DNA, voltage, and the optimal electrophoresis condition were evaluated. The result showed the PCR reaction procedure with a total volume of ۲۰ μL with ۶۰ ng template DNA was as follows: initiation at ۹۴ ℃ for ۴ min, denaturation at ۹۴ ℃ for ۱ min, annealing at ۴۸.۰ ℃~۵۴.۵ ℃ for ۵۰ seconds (annealing temperature depend on different primer), extension at ۷۲ ℃ for ۱.۵ min, ۳۰-۳۹ cycles, final extension at ۷۲ ℃ for ۱۰ min. Also, optimal electrophoresis condition was determined on ۱.۵% agarose gel at ۹۸ v for one hour.

Keywords:

Genetic diversity , Molecular marker , Optimal parameters