2.3. RNA Extraction, cDNA Library Construction/Preparation

The total RNA was isolated from 50 mg tail tissue using the TRIzol reagent, as reported previously by Ranadive et al. (2018) [4]. The quality and quantity of the isolated RNA were estimated using Qubit Fluorimeter 3.0 and Agilent Tapestation 2200. Then, 1 μg of isolated RNA was used to prepare the transcriptome library using Truseq RNA Library prep kit V2 (Illumina Inc., San Diego, CA, USA, Cat# RS-122-2001) as per the manufacturer’s protocol. The libraries were sequenced on Illumina HiSeq 2500 platform in 100 bp paired end sequencing.