Sharma Davinder, Golla Naresh, Singh Dheer, Onteru Suneel K
Molecular Endocrinology, Functional Genomics, and Systems Biology Lab, Animal Biochemistry Division, ICAR-National Dairy Research Institute, Karnal, India.
J Cell Physiol. 2018 Mar;233(3):1971-1974. doi: 10.1002/jcp.25951. Epub 2017 Jun 5.
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.
基于新一代测序（NGS）的RNA测序（RNA-Seq）和转录组分析为揭示复杂的生物学过程提供了契机。成功的RNA-Seq和转录组分析需要大量高质量的RNA。然而，从脂质含量高且细胞数量少的顽固性脂肪组织（AT）中分离出NGS质量的RNA极其困难。此外，AT脂质的数量和生化组成因动物物种而异，这可能对RNA提取产生不同程度的抗性。目前可用的方法在一个物种中可能有效，但在另一个物种中可能几乎无效。在此，我们报告了一种两步法方案，用于从广泛的动物物种的AT中提取NGS质量的RNA。
