GSM5665226: IMPA 2; Capra hircus; RNA-Seq
Extraction of total RNA from sequenced samples using Trizol reagent (Takara, Dalian, China) The lncRNA library is constructed using a chain-specific library. The method for synthesizing the first strand of cDNA by reverse transcription is the same as the normal method of NEB library construction. The difference is that when the second strand is synthesized, dTTP in dNTPs is replaced by dUTP. After that, cDNA end repair, A-tailing, ligation of sequencing adapters, and length screening were also performed, and then the second strand of cDNA containing U was degraded by USER enzyme, and then PCR amplification was performed to obtain a library. Finally, twenty libraries were sequenced at Novogene Co. Ltd. (Beijing, China) on Illumina HiSeq Sequencing System.