华北农学报2011，Vol.26Issue(5)：92-96,5.
牡丹SCoT分子标记正交优化及引物筛选
Orthogonal Optimization of SCoT-PCR System and Primer Screening of Tree Peony
摘要
Abstract
The orthogonal design table of L16(4 )was used to optimize the five factors( Taq DNA polymerase, Mg2+ , DNA template, dNTPs and primer concentrations) of SCoT-PCR( start codon targeted polymorphism) system in tree peony. The results showed that the optimized system was as follows; a total volume of 20 μL including 1. 0 ng/μL DNA template,0. 50 U Taq polymerase,0. 60 μmol/L primer,2. 50 mmol/L Mg2+ ,0. 25 mmol/L dNTPs and 1 PCR Buffer. Each factor had different effect on the results. The concentration of Mg * was the key factor affecting the SCoT-PCR system. The optimized SCoT-PCR system was tested on seventeen peony genus,and the result was stable and reliable. From the 36 primer combinations tested,Twenty-four were selected with clear band patterns and abundant polymorphism. The optimized SCoT-PCR system and polymorphism primmer combinations provided the basis for further research on Paeonia suffruticosa.关键词
牡丹/SCoT-PCR/反应体系/正交设计/引物筛选Key words
SCoT-PCR/ Reaction system/Orthognal design/Primers screening分类
农业科技引用本文复制引用
侯小改,王娟,贾甜,张钰乾,侯娟,李嘉珏..牡丹SCoT分子标记正交优化及引物筛选[J].华北农学报,2011,26(5):92-96,5.基金项目
国家自然科学基金(31070620) （31070620）
河南省重大科技攻关项目(091100110100) （091100110100）
河南省重点科技攻关项目(092102110024) （092102110024）
洛阳市科技攻关项目(0802017A) （0802017A）
