Establishment and Optimization of ISSR-PCR System for Dendrocalamus minor var.amoenus
QU Yinquan
XU Wen
SHEN Lu
HE Tianyou
SHI Chengkun
RONG Jundong
CHEN Liguang
ZHENG Yushan
Abstract：The purpose of this study was to establish optimized ISSR system for Dendrocalamus minor var.amoenus.The single factor design was applied for optimizing five factors in ISSR amplification system,such as Mg2 +,dNTPs,Taq DNA polymerase,primer and template DNA concentration.The results showed that the optimal 20 μL ISSR-PCR reaction system for Dendrocalamus minor var.amoenus contained 3.0 mmol/L Mg2+,0.20 mmol/L dNTPs,1.25 U Taq DNA polymerase,0.6 μmol/L primer,10 ng DNA template,2 μL 10 × Buffer,8.55 μL ddH20.The amplification procedure was as follows:94 ℃ 5 min;94 ℃ 45 s,52.7 ℃30 s,72 ℃90 s,40 cycles;72 ℃ 10 main,4 ℃ for storage.Under this optimal ISSR amplification system,16 primers were screened with high polymorphism from 100 primers.The test showed that the system was stable,reliable and applicative.
Keywords：Dendrocalamus minor var.amoenusISSRreaction systemoptimization
Publication Date：2017-07-02
Online Publishing Date：2026-09-12(First online date of this platform, not the publication date of the document)
Pages：6( 86-91 )