WANG Qian-chun,HUANG Xia,LI Guang-sen.Establishment of the SRAP-PCR Reaction System of Pinus contorta[J].Northern Horticulture,2013,37(02):83-86.
小干松SRAP-PCR反应体系的建立
- Title:
- Establishment of the SRAP-PCR Reaction System of Pinus contorta
- 文章编号:
- 1001-0009(2013)02-0083-04
- Keywords:
- Pinus contorta; SRAP; reaction system
- 分类号:
- S 791.256
- 文献标志码:
- A
- 摘要:
- 以小干松针叶基因组DNA为模板，采用L16(45)正交实验设计，对SRAP-PCR反应体系中的Taq酶、Mg2+、dNTPs、模板DNA和引物5个因素在4个水平上进行优化。结果表明：小干松SRAP-PCR 20 μL反应体系最佳组合为：Taq酶0.5 U，Mg2+浓度2.5 mmol/L，dNTPs浓度0.15 mmol/L，模板DNA含量60 ng，引物0.2 μmol/L。使用12对SRAP引物，采用优化后的体系进行SRAP-PCR反应，表明优化的体系很好地满足了小干松基因组DNA进行SRAP的扩增要求。
- Abstract:
- To establish the optimal SRAP reaction system of Pinus contorta，its genomic DNA was extracted as template and orthogonal experimental design L16(45) were used.Four levels of five factors (Taq polymerase，Mg2+，dNTPs，DNA template and primer) were optimised.The results showed that according to the amplification results，an optimal and stable SRAP-PCR system for Pinus contorta was 0.5 U Taq polymerase，2.5 mmol/L Mg2+，0.15 mmol/L dNTPs，60 ng DNA template and 0.2 μmol/L primer in total 20 μL reaction solution.12 pairs SRAP primer were used in this optimized SRAP-PCR system，the results showed that this system could meet the requirement for SRAP amplification of Pinus contorta.
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备注/Memo
第一作者简介：王骞春（1979-），男，硕士，工程师，现主要从事林木遗传育种等研究工作。