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Rapid and Efficient Method of Total RNA Isolation from Milk Fat for Transcriptome Analysis of Mammary Gland

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Proceedings of the National Academy of Sciences, India Section B: Biological Sciences Aims and scope Submit manuscript

Abstract

Isolation of good quality ribonucleic acid (RNA) from mammary glands of elite animals is often hindered by invasive method of mammary tissue sampling, ethical permission, time consuming, and repeated biopsies from the same animal at different time points. The aim of the present study was to optimize a protocol for RNA isolation from milk fat that is suitable for transcriptome analysis. In this study, authors isolated good quality RNA from milk fat of goats and buffalo milk by combining two methods namely Trizol and GenElute mammalian RNA isolation kit. The results showed good quality as well as high quantity of total RNA suitable for downstream applications like reverse transcriptase-quantitative polymerase chain reaction and next generation sequencing.

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Acknowledgements

The authors would like to thank Dr. Devendra Pathak, Department of Veterinary Anatomy, for assisting in making of paraffin embedded tissue debris after Trizol digestion of milk fat. They thank Department of LPM, GADVASU, especially to Dr. Saini, Professor and Head and to Dr. Sandeep Kaswan for allowing and helping them to collect milk samples from goats. The authors also thank Dr. Puneet Malhotra, Department of AGB, for allowing them to collect buffalo milk from the farm.

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Authors and Affiliations

  1. School of Animal Biotechnology, Guru Angad Dev Veterinary and Animal Science University, Ludhiana, Punjab, 141004, India

    Shanti Choudhary & Ratan K. Choudhary

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  1. Shanti Choudhary
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  2. Ratan K. Choudhary
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Correspondence to Ratan K. Choudhary.

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The authors declare no any competing interests for publishing this manuscript.

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Significance statement This paper reports a non-invasive method of RNA isolation from milk fat of goat and buffalo that is suitable for global gene expression analysis. This method would not be impeded by ethical concerns for obtaining tissue biopsy and repeated sampling. Utility of this protocol could be extended to other species for transcriptome analysis without undergoing invasive mammary tissue biopsy.

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Choudhary, S., Choudhary, R.K. Rapid and Efficient Method of Total RNA Isolation from Milk Fat for Transcriptome Analysis of Mammary Gland. Proc. Natl. Acad. Sci., India, Sect. B Biol. Sci. 89, 455–460 (2019). https://doi.org/10.1007/s40011-017-0955-8

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  • DOI: https://doi.org/10.1007/s40011-017-0955-8

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