A highly efficient method for extracting next‐generation sequencing quality RNA from adipose tissue of recalcitrant animal species. Issue 3 (5th June 2017)

• Record Type: Journal Article
• Authors: Sharma, Davinder; Golla, Naresh; Singh, Dheer; Onteru, Suneel K.
• Is Part Of: Journal of cellular physiology. Volume 233:Issue 3(2018:Mar.)
• Page Start: 1971
• Page End: 1974
• Publication Date: 2017-06-05
• Subjects: adipose tissue -- next‐generation sequencing -- recalcitrant animal species -- RNA isolation; Physiology -- Periodicals; Cell physiology -- Periodicals; 571.6
• DOI: 10.1002/jcp.25951
• Languages: English
• ISSNs: 0021-9541

Abstract

Abstract : The next‐generation sequencing (NGS) based RNA sequencing (RNA‐Seq) and transcriptome profiling offers an opportunity to unveil complex biological processes. Successful RNA‐Seq and transcriptome profiling requires a large amount of high‐quality RNA. However, NGS‐quality RNA isolation is extremely difficult from recalcitrant adipose tissue (AT) with high lipid content and low cell numbers. Further, the amount and biochemical composition of AT lipid varies depending upon the animal species which can pose different degree of resistance to RNA extraction. Currently available approaches may work effectively in one species but can be almost unproductive in another species. Herein, we report a two step protocol for the extraction of NGS quality RNA from AT across a broad range of animal species. Abstract : NGS‐quality RNA isolation is extremely difficult from recalcitrant adipose tissue (AT) with high lipid content and low cell numbers. Currently available approaches yield variable and unsatisfactory results with AT samples.A highly efficient novel protocol was developed for the extraction of NGS quality RNA from AT across a broad range of animal species.