适用于微卫星标记的湿地松、加勒比松dna快速提取法 - 生物技术通报.pdfVIP

生物技术通报 BIO TE CHN OLOG Y B ULLETIN 20 10 1 DNA 李义良 赵奋成 张应中 吴惠姗 (, 510520) : 以湿地松 ( P inus elliottii, PEE ), 包括古巴加勒比松 (P caribaea varcaribaea, PCC)洪都拉斯加勒比松 (P caribaea varhondurensis, PCH )巴哈马加勒比松(P caribaea varbahm aensis, PCB) 3个变 在内的加勒比松, 侧枝顶芽为试 验材料, 使用 RETSCH MM 400混合球磨仪破碎植物组织, 然后利用改良 CTAB法提取基因组 DNA, 完成 48个样品仅用 1 h, 1 个工作日可提取 300个样品以上, DNA 分子量大于 20 kb, 03 g样品可提取 DNA 20- 0 g, 能够满足几百次 PCR反应; 利用 所提 DNA进行 SSR分析, 条带清晰, 多态性好, 说明该方法提取的 DNA 完全可以满足 SSR 反应的需要本研究为 SSR标记 用于湿地松加勒比松分子标记辅助选择育 提供了经济高效可靠的 DNA 提取方法 : 湿地松 加勒比松 DNA提取 SSR分子标记 Study of Rapid DNA Extraction M ethod Suitable for SSR Analysis in Slash P ine and Caribbean P ine LiY iliang Zhao Fencheng Zhang Y ingzhong W u Huishan ( Institute of Forestry, Guangdong Academy of Forestry, Guangzhou 510520) A bstrac:t One mi proved CTAB method to extracted genom ic DNA from term inal buds on lateral branch of slash pine and Caribbe an pine was providedAccord ing to the mi proved m ethod and the use ofRETSCH MM 400 fragm ented plant tissue for extracted ofDNA, the efficiencyw as raised compared to the norm al processIn the expermi ent, 48 sam ples cou ld be treated one tmi e and the process could be fin ished in one hour, and DNA of 300 sam ples could be ob tained in one w orkdayIt w as estmi ated that m olecu lar weight of the ex tracted DNA is larger than 20 kbAnd 20- 0 g DNA cou ld be extracted from 03 g term inal bud, which wou ld be enough for hundreds of tmi es of PCR reactionThe extracted DNA w as using in SSR analysis, the band ing patterns w as clear and w ith good polym orph ism, and the quality of DNA extracted using thism ethodwas suitable for SSR analys isThis study provide an econom ic, efficient, and reliab le