Welcome to Chinese Agricultural Science Bulletin,

Chinese Agricultural Science Bulletin ›› 2014, Vol. 30 ›› Issue (10): 177-181.doi: 10.11924/j.issn.1000-6850.2013-2221

Special Issue: 园艺

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Establishment and Optimization of ISSR Reaction System for Dendrobium

  

  • Received:2013-08-19 Revised:2013-09-24 Online:2014-04-05 Published:2014-04-05

Abstract: The research aimed to establish and optimize a stable ISSR-PCR reaction system for Dendrobium. In this paper, the key factors of ISSR-PCR reaction system including DNA template concentration, dNTP, 10×PCR Buffer, primer concentration, DNA Taq polymorphism were investigated and optimized. The stable reaction system could amplify high levels of polymorphism, good repeatability and clear band patterns. ISSR system (total volume of 20μL) established was as follows: template DNA 10 ng/μL 3μL, dNTP 2.5 mmol/L 1.5μL, 10×PCR Buffer 3.0μL, primer 4μmol/L 3.5μL, Taq polymorphism 5 U 0.2μL. It showed that the optimized system in this experiment could be applied in the ISSR analysis for Dendrobium. This research provided technical support for genetic diversity and genetic relationship research, linkage map construction, QTL location, gene mapping and gene cloning and so on.