Optimization for RAPD Reaction System of Armeniaca sibirica

Abstract

Abstract: The factors influencing RAPD analysis, including Taq polymerase, Mg2+, dNTP, primers, template DNA’s concentration, denaturing time and thermal cycles in Armeniaca. sibirica (L.) Lam. were studied. An optimal PCR system for RAPD in fresh leaves of A. sibirica (L.) Lam. has been found： in 20μL reaction solution, contained 1U Taq polymerase, 2.0mMMg2+, 0.1mM dNTP, 0.5μM primers, 50ng template DNA. The amplification program was devised： initial denaturing at 94℃ for 4mins; denaturing at 94℃ for 1mins, annealing at 36℃ for 1min, extension at 72℃ for 1.5mins, 10cycles; denaturing at 94℃ for 30s, annealing at 36℃ for 40s, elongation at 72℃ for 60s, 30 cycles; final extension at 72℃ for 5mins.

CLC Number:

S662.2

Li Zhenjiang, Ge Huibo,, Zhang Xueying, Wang Yong. Optimization for RAPD Reaction System of Armeniaca sibirica[J]. Chinese Agricultural Science Bulletin, 2007, 23(6): 169-169.

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