Development and Optimization of Multiplex PCR-Based Marker of Ms Locus in Onion
Wang Zhenbao
Huo Yumeng
Liu Bingjiang
Miao Jun
Yang Yanyan
Gao Limin
Kong Suping
Cheng Fei
Chen Ning
Wu Xiong
Abstract：Through designing and screening primers on the basis of flanking sequence of Ms locus, a compatible multiplex PCR-based marker ( MK4 ) was obtained , and its reaction system and procedure were optimized.The optimal PCR reaction system consisted of 2.5μL of 10 ×PCR buffer (Mg2+free), 4μL of 25 mmol/L MgCl2, 6 μL of 2.5 mmol/L dNTP, 1 μL of DNA template (about 50 ng), 1 μL of each of 10 mmol/L primer, 0.6μL of 5 U/μL rTaq DNA polymerase , and finally adding ddH 2 O to 25μL.The PCR re-action procedure was initial denaturing at 94℃ for 5 min, followed by 35 cycles of denaturing at 94℃ for 30 s, annealing at 65.4℃for 45 s, extending at 72℃for 1 min, and then extending at 72℃for 10 min in the end.Three genotypes ( MsMs, Msms, msms) of Ms locus could be clearly distinguished by MK 4 based on the optimal reaction conditions and procedure .The reaction was simple and had good stability .
Keywords：OnionMale sterilityMs locusMultiplex PCR-based marker
Publication Date：2014-01-01
Online Publishing Date：2026-05-22(First online date of this platform, not the publication date of the document)
Pages：5( 7-11 )