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Extraction of high-molecular-weight DNA from dry root tissue ofBerberis lycium suitable for RAPD

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Abstract

A simple protocol for DNA isolation from dry roots ofBerberis lycium is described. Four-year-old dry roots are used, and the isolated DNA is suitable for analysis by means of restriction enzyme digestion and random amplification of polymorphic DNA (RAPD). The method involves a modified CTAB procedure using 1% PVP to remove polysaccharides and purification using low-melting-temperature agarose. DNA is amplified by means of PCR using 10-mer random primers from Operon Biotechnologies, Inc. (USA), and DNA samples are digested withTaq I,Hind III andEcoR I and examined on agarose gels. The RAPD reaction is performed according to the 1990 protocol by Williams et al.

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Authors and Affiliations

  1. Molecular Taxonomy Laboratory, Molecular Biology and Genetic Engineering Division, National Botanical Research Institute, 226001, Lucknow, UP, India

    Anil Kumar & P. Pushpangadan

  2. Pharmacognosy and Ethnopharmacology Division, National Botanical Research Institute, 226001, Lucknow, UP, India

    S. Mehrotra

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  1. Anil Kumar
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  2. P. Pushpangadan
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  3. S. Mehrotra
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Correspondence to Anil Kumar.

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Kumar, A., Pushpangadan, P. & Mehrotra, S. Extraction of high-molecular-weight DNA from dry root tissue ofBerberis lycium suitable for RAPD. Plant Mol Biol Rep 21, 309 (2003). https://doi.org/10.1007/BF02772807

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  • DOI: https://doi.org/10.1007/BF02772807

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