新疆野苹果基因组提取优化及模板浓度对ISSR-PCR影响

Journal of Tianjin Agricultural University ›› 2017, Vol. 24 ›› Issue (1): 15-18.

• Researches and Scientific Notes • Previous Articles Next Articles

Optimization of Genome Extraction and Effect of Template Concentration on ISSR-PCR in Malus sieversii

ZHANG Yun-xiu¹, YANG Mei-ling², YU Wei-wei¹, LI Fang¹, LIU Bin¹, LONG Hong¹, YAN Guo-rong^{1,Corresponding Author}

1. College of Horticulture and Landscape, Tianjin Agricultural University, Tianjin 300384, China;
2. College of Life Sciences,Nankai University, Tianjin 300071, China

Received:2016-05-13 Online:2017-03-31 Published:2019-10-15

Abstract

Abstract: As Malus sieversii dried leaves for materials, compared and analyzed the result of extracting genomic DNA with the CTAB method and four kinds of modified CTAB method, besides, to determine the best template concentration for ISSR- PCR. Results showed that the purity and quantity of DNA with five methods has much difference, grinding with PVP can effectively remove polyphenols; adding1/10 volume CTAB cracking in the supernatant fluid （0.7 mol/L NaCl, 10% CTAB） can effectively remove polysaccharide; isopropyl alcohol precipitating with 1/9 volume sodium acetate can get white precipitation quickly . As for the M. sieversii ISSR amplification system study, found that ISSR-PCR amplified result is best with DNA template concentration in 400 ~ 1 000 ng/μL. DNA extraction and the concentration of template optimization provide theoretical basis for subsequent molecular biology research.

Key words: Malus sieversii, DNA extraction, template concentration, optimization

CLC Number:

Q946-33

ZHANG Yun-xiu, YANG Mei-ling, YU Wei-wei, LI Fang, LIU Bin, LONG Hong, YAN Guo-rong. Optimization of Genome Extraction and Effect of Template Concentration on ISSR-PCR in Malus sieversii[J]. Journal of Tianjin Agricultural University, 2017, 24(1): 15-18.

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Optimization of Genome Extraction and Effect of Template Concentration on ISSR-PCR in Malus sieversii

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