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1 January 2012 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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Abstract

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 (1X 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 µg/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.

Sayantani Das, Maumita Bandyopadhyay, and Subir Bera "Optimization of Protocol for Isolation of Genomic DNA from Leaves of Selaginella Species Suitable for RAPD Analysis and Study of their Genetic Variation," American Fern Journal 102(1), 47-54, (1 January 2012). https://doi.org/10.1640/0002-8444-102.1.47
Published: 1 January 2012
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