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A Simple Method for the Efficient Isolation of Genomic DNA from Lactobacilli Isolated from Traditional Indian Fermented Milk (dahi)

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Abstract

A simple, inexpensive and effective genomic DNA isolation procedure for Lactobacillus isolates from traditional Indian fermented milk (dahi) is described. A total of 269 Lactobacillus isolates from fermented milk collected from four places in North and west India were tested for lysis by an initial weakening of the Gram positive cell wall with Ampicillin followed by Lysozyme treatment. The average genomic DNA yield was ~50 μg/ml log phase culture. Quality and repeatability of the method was found to be adequate for subsequent molecular applications. The quality of the genomic DNA isolated by this method was verified by restriction digestion and polymerase chain reaction (PCR). No inhibition was observed in subsequent PCR amplification and restriction digestion. The presented method is rapid, cheap and useful for routine DNA isolation from gram positive bacteria such as Lactobacillus.

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Acknowledgments

This work was supported by a grant for the “Microbial Diversity and Identification Project” funded by the National Bureau of Agriculturally Important Organisms, Mau, Uttar Pradesh, India. The authors would like to thank Dr. D. K. Arora (Director, NBAIM) for the help in supporting this work.

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Authors and Affiliations

  1. Animal Biotechnology Center, National Dairy Research Institute, Karnal, India

    Sachinandan De, Gurpreet Kaur, Amit Roy, Ramakant Kaushik, Paras Yadav, Tirtha Kumar Datta & Surender Lal Goswami

  2. Dairy Microbiology Division, National Dairy Research Institute, Karnal, India

    Gaurav Dogra & Rameshwar Singh

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  1. Sachinandan De
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  2. Gurpreet Kaur
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  3. Amit Roy
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  4. Gaurav Dogra
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  9. Surender Lal Goswami
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Correspondence to Sachinandan De.

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De, S., Kaur, G., Roy, A. et al. A Simple Method for the Efficient Isolation of Genomic DNA from Lactobacilli Isolated from Traditional Indian Fermented Milk (dahi). Indian J Microbiol 50, 412–418 (2010). https://doi.org/10.1007/s12088-011-0079-4

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  • DOI: https://doi.org/10.1007/s12088-011-0079-4

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