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Journal of Agriculture ›› 2016, Vol. 6 ›› Issue (3): 46-50.doi: 10.11923/j.issn.2095-4050.cjas15080012

Special Issue: 园艺

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Optimization and Verification of ISSR-PCR Reaction System for Lycopersicon esculentum

  

  • Received:2015-08-20 Revised:2015-11-16 Accepted:2015-11-25 Online:2016-03-28 Published:2016-03-28

Abstract: To study the optimization of ISSR-PCR reaction system for Lycopersicon esculentum, the effects of five main factors (TaqDNA polymerase dosage, dNTP concentration, primer concentration, DNA templates concentration and annealing temperature of each primer) on ISSR amplification were investigated, to lay a foundation for ISSR technique in tomato molecular assisted breeding. The results showed that the optimal ISSR-PCR reaction system (20 μL) contained 50 ng DNA template, 0.3 μmol/L primer, 0.5 mmol/L dNTP, 2 U TaqDNA polymerase, and 2.0 μL 10×PCR buffer. In addition, the optimal annealing temperature of each primer for ISSR-PCR reaction was different.