Optimization of Genomic-SSR Reaction System in Liriodendron
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1.
Key Laboratory of Forest Genetics & Biotechnology of Ministry of Education, Nanjing Forestry University, Nanjing 210037, Jiangsu, China
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2.
The National Engineering Laboratory for Breeding of Endangered Medicinal Materials, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100193, China
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Received Date:
2013-02-08
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Abstract
The optimization of the Genomic-SSR reaction system is a basic protocol when the Genomic-SSR is used for genetic analysis in Liriodendron. The concentrations of Mg2+, dNTP, Primer and rTaq were tested by L9(34) orthogonal experiment and single factor gradient experiment to gain the optimal reaction system. The results indicated that the optimal reaction system should contain 75 ng of genomic DNA, 1 μL of 10×buffer, 0.4 μL of 10 mmol·L-1 dNTP, 0.75 μL of 2.5 mmol·L-1 MgCl2, 0.25μL of 10 μmol·L-1 Primer, 0.05 μL of 5 U rTaq and final volume of 10μL. Repeated trials and two verification tests showed that this optimal reaction system was stable, reliable, efficient and suitable for the applications of Genomic-SSR in Liriodendron population genetics and quantitative genetics research.
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