安徽农业大学学报2012，Vol.39Issue(3)：412-416,5.
多花黄精ISSR反应体系的建立及正交优化设计
Establishment and optimization of an ISSR reaction system for Polygonatum cyrtonema Hua using orthogonal design
摘要
Abstract
In this paper, the orthogonal design of five factors (Tag DNA polymerase, buffer (Mg2+), DNA template, dNTP, and primer) in four levels was used to optimize the ISSR-PCR amplification system on Polygonatum sibiricum. Finally, a satisfactory ISSR (Inter Simple Sequence Repeat) reaction system for Polygonatum sibiricum with desirable repeatability and polymorphic bands was established. It contained 1.0 U Taq DNA polymerase, 3.5 mmolL"' buffer (Mg2*), 80 ng DNA template, 0.08 mmolL'1 dNTP and 0.16 umol-U1 primer respectively in a 25 uL ISSR reaction system. The optimal annealing temperature of every prime for ISSR-PCR reaction was proposed by gradient PCR with established ISSR reaction, the optimal annealing temperature was 54.8°C for primer UBC841, and different primers had different annealing temperatures. This optimized ISSR reaction system would provide the basis for the analysis of classification, map construction and gene localization of Polygonatum sibiricum.关键词
多花黄精/ISSR/正交设计/反应体系/优化/退火温度Key words
Polygonatum cyrtonema Hua/ ISSR/ orthogonal design/ reaction system/ optimization/ annealing temperature分类
农业科技引用本文复制引用
张红梅,邵元华,龙甜甜,项艳,王进,汤锋..多花黄精ISSR反应体系的建立及正交优化设计[J].安徽农业大学学报,2012,39(3):412-416,5.基金项目
国家林业公益性行业科研专项(201004055)资助. （201004055）
