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Excised radicle tips as a source of genomic DNA for PCR-based genotyping and melting curve analysis in cotton

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Abstract

Genomic DNA isolation in cotton is complicated because of the presence of secondary metabolites that are inhibitory to PCR amplification. We report here that radicle tips, but not other parts of cotton seedlings, yield high-quality DNA that is readily amenable for PCR. The radicle-tip-excised seedlings retain viability because of the formation of adventitious roots. We demonstrate the utility of this method in distinguishing homozygotes from heterozygotes in a cotton breeding population and in hybrid seed purity testing.

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Abbreviations

Bt:

Bacillus thuringiensis

HRMA:

high-resolution melting curve analysis

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Acknowledgements

The first author acknowledges the financial assistance of Department of Biotechnology (DBT), India, through its post-doctoral program.

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Author notes

  1. P Srinivasa Rao

    Present address: International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Patancheru, Hyderabad, 502 324, India

Authors and Affiliations

  1. Centre for Cellular and Molecular Biology (CCMB), Uppal Road, Hyderabad, 500 007, India

    P Srinivasa Rao & Ramesh V Sonti

  2. Nuziveedu Seeds, Road No. 12, Banjara Hills, Hyderabad, 500 034, India

    P Sateesh Kumar

Authors
  1. P Srinivasa Rao
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  2. P Sateesh Kumar
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  3. Ramesh V Sonti
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Corresponding author

Correspondence to Ramesh V Sonti.

Additional information

MS received 12 March 2012; accepted 10 October 2012

Corresponding editor: Utpal Nath

[Rao PS Kumar PS and Sonti RV 2013 Excised radicle tips as a source of genomic DNA for PCR-based genotyping and melting curve analysis in cotton. J. Biosci. 38 1–6] DOI10.1007/s12038-012-9285-4

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Rao, P.S., Kumar, P.S. & Sonti, R.V. Excised radicle tips as a source of genomic DNA for PCR-based genotyping and melting curve analysis in cotton. J Biosci 38, 167–172 (2013). https://doi.org/10.1007/s12038-012-9285-4

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  • DOI: https://doi.org/10.1007/s12038-012-9285-4

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