Optimization on multiplex PCR amplification system of SSR marker forauthenticity and purity identification of maize varieties

Abstract

Abstract: In order to establish a stable and reliable identification of maize authenticity and purity of the SSR markers, genetic DNA extraction, SSR primers and multiplex PCR reaction procedures were optimized. The results showed that the high purity and good integrity of DNA could be obtained using above 75℃ preheating mortar and 95℃ of 1.5×CTAB extraction buffer to grind material. Redesign and optimize the primer sequence using the software Primer Premier 5.0 and Oligo 6.72, eight sets of multiplex PCR amplified reactions, which was consisting of 21 pairs of universal SSR primers and a three-step procedure, were with uniform procedures, no intersection among the amplified fragments, amplified bands clear and stable amplification results. The testing efficiency of the amplification system increased 2.6 times higher than a single pair of primers SSR.

CLC Number:

S513.01

CHANG Hong, WANG Han-ning, ZHANG Jin-wen, WANG Wei,
LU Ze-quan, LI Yin, WANG Di. Optimization on multiplex PCR amplification system of SSR marker forauthenticity and purity identification of maize varieties[J]. Acta Prataculturae Sinica, 2010, 19(2): 204-211.

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