浙江农林大学学报2012，Vol.29Issue(6)：960-965,6.
光皮桦EST-SSR PCR反应体系的优化
Optimization of EST-SSR PCR reaction system for Betula luminifera
摘要
Abstract
To obtain stable PCR product, providing the basis for further study, orthogonal design was used to optimize an Expressed Sequence Tags (EST)-Simple Sequence Repeat (SSR) Polymerase Chain Reaction (PCR) amplification system for Betula luminifera in terms of five factors containing rTaq DNA polymerase, Mg2+, DNA template, dNTPs and primers in five levels. Results showed an optimal EST-SSR PCR system for B. luminifera was 4 mg·L-1 DNA template, 0.500 μmol·L-1 primers, 1.250 mmol·L-1 Mg2+, 0.250 mmol·L-1 dNTPs, and 0.072 5×16.67 mkat·L-1 rTaq DNA polymerase. And the optimal annealing temperature to each primers for SSR PCR reaction system is determined by gradient PCR. Using the optimal reaction system, stable and clear polymorphic bands were acquired. [Ch, 6 fig. 3 tab. 15 ref.]关键词
林木育种学/光皮桦/EST-SSR/PCR反应体系/正交设计Key words
forest tree breeding/Betula luminifera/EST-SSR/PCR reaction system/Orthogonal design分类
农业科技引用本文复制引用
陈争,姜小凤,童再康..光皮桦EST-SSR PCR反应体系的优化[J].浙江农林大学学报,2012,29(6):960-965,6.基金项目
浙江省重大科技专项(2008C02004-1) （2008C02004-1）
