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首页> 外文期刊>American Fern Journal >Optimization of Protocol for Isolation of Genomic DNA from Leaves of Selaginella Species Suitable for RAPD Analysis and Study of their Genetic Variation
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Optimization of Protocol for Isolation of Genomic DNA from Leaves of Selaginella Species Suitable for RAPD Analysis and Study of their Genetic Variation

机译：适于RAPD分析的卷柏属植物叶片基因组DNA分离方法的优化及其遗传变异研究

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摘要

A simple and efficient protocol for isolating genomic DNA from leaves of Selaginella spp. (S. delicatula, S. repanda, S. bryopteris, S. plana, S. monospora) was developed, involving a modified CTAB protocol of Rogers and Benedich (1994). Increasing the incubation time with the precipitation buffer (IX CTAB) from 1-3 hours to 12-14 hours helped achieve higher quantity genomic DNA from the specimens, when compared with DNA extracted by protocols reported by Dellaporta et al. (1983), Murray and Thompson (1980) and Doyle and Doyle (1987). The DNA yield ranged from 846-1836 ng/ml from fresh and herbaria-preserved leaf samples. The DNA samples were found suitable for genetic diversity analysis with Random Amplified Polymorphic DNA (RAPD) markers. Nine random primers (OPA A17, OPB 4, OPB13, OPC 2, OPC 11, OPD 5, OPG 2, OPG 19 and OPK 10) were studied, of which two primers (OPD 5 and OPG 2) yielded reproducible amplification profile of polymorphic fragments.
机译：从卷柏属植物的叶片中分离基因组DNA的简单有效的协议。 （S. delicatula，S。repanda，S。bryopteris，S。Plana，S。monospora）已开发，涉及Rogers和Benedich（1994）的改良CTAB方案。与通过Dellaporta等人报道的方案提取的DNA相比，用沉淀缓冲液（IX CTAB）的孵育时间从1-3小时增加到12-14小时有助于从样品中获得更高数量的基因组DNA。 （1983），默里和汤普森（1980）和道尔和道尔（1987）。新鲜和保留草叶的DNA样品的DNA产量范围为846-1836 ng / ml。发现该DNA样品适用于使用随机扩增多态性DNA（RAPD）标记进行遗传多样性分析。研究了9种随机引物（OPA A17，OPB 4，OPB13，OPC 2，OPC 11，OPD 5，OPG 2，OPG 19和OPK 10），其中两种引物（OPD 5和OPG 2）产生了可复制的多态性扩增图谱碎片。

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  • 来源
    《American Fern Journal》 |2012年第1期|p.47-54|共8页
  • 作者

    Sayantani Das; Maumita Bandyopadhyay; Subir Bera;

  • 作者单位

    Centre of Advanced Studies, Department of Botany, University of Calcutta, 35, B.C.Road, Kolkata- 700019, India;

    Centre of Advanced Studies, Department of Botany, University of Calcutta, 35, B.C.Road, Kolkata- 700019, India;

    Centre of Advanced Studies, Department of Botany, University of Calcutta, 35, B.C.Road, Kolkata- 700019, India;

  • 收录信息 美国《科学引文索引》(SCI);
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    DNA extraction; RAPD; selaginella; modified protocol;

    机译：DNA提取;RAPD;卷柏修改协议;

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