Optimization of ISSR Reaction System of Hemp and Elementary Screening of Primers

doi:10.11924/j.issn.1000-6850.2013-2549

Chinese Agricultural Science Bulletin ›› 2014, Vol. 30 ›› Issue (12): 105-109.doi: 10.11924/j.issn.1000-6850.2013-2549

Optimization of ISSR Reaction System of Hemp and Elementary Screening of Primers

Received:2013-09-27 Revised:2013-11-04 Online:2014-04-25 Published:2014-04-25

Abstract

Abstract: The five factors affected ISSR reaction, including dNTP, TaqDNA polymerse, concentrations of primer, predegeneration time, annealing temperature etc., were tested to optimize ISSR amplification system by single factor experiment method for finding genetic diversity of hemp. The results showed that: the optimum reaction system in 25μL volume was dNTP 200μmol/L, template 1 ng/μL, primer 200 nmol/L, Taq DNA polymerase 1 U (2.5 pmol/μL), predegeneration time 3 min (94℃), annealing temperature 48℃. Moreover, 8 primers in 30 primers were screened out through the above stable reaction system. The repeatable and stable system was completely suitable to analyze the hemp genetic information.

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Optimization of ISSR Reaction System of Hemp and Elementary Screening of Primers

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