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《Journal of Anhui Agricultural Sciences》 2008-29
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Establishment of DNA Extraction and ISSR-PCR Reacting System of Tutcheria kweichowensis

ZHAO Yang et al (College of Forestry,Guizhou University,Guiyang,Guizhou 550225)  
[Objective] The research aimed to lay the foundation for the genetic diversity analysis of Tutcheria kweichowensis by ISSR technology.[Method] The materials of the fresh leaves,dry leaves and seeds of Tutcheria kweichowensis and the improved methods of SDS and CTAB were used for the DNA extraction experiment,and the concentration and purity of the extracted DNA were determined.Through single-factor gradient test,the optimized ISSR-PCR reaction system was screened out.[Result] When extracting the leaf DNA of Tutcheria kweichowensis with the methods of CTAB and SDS,there were small differences in quality and density between the fresh leaves and dry leaves.There were higher density and more protein residues by CTAB than those by SDS.The quantity of DNA with SDS was a little less than that of CTAB but with the highest quality.Both the quality and density of DNA extracted from seeds were worse,but still qualified for ISSR molecular marking experiment.The appropriate ISSR-PCR reacting conditions were as follows: 20 ul reacting system,10×Buffer,1 U Taq DNA enzyme,0.2 μmol/L primer,0.25 mmol/L L dNTP and 40 ng template DNA.[Conclusion] This ISSR reaction system was suitable for the genetic diversity analysis of Tutcheria kweichowensis.
【Fund】: 贵州大学引进人才基金项目(X060055)
【CateGory Index】: Q943.2
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