GSM5548047: Control 6; Mus musculus; RNA-Seq
RNA was extracted using Qiagen Rneasy kit. After extracting the total RNA from the tissues, mRNA is then enriched by using the oligo (dT) magnetic beads from the total RNA. By using the fragmentation buffer, the mRNA is fragmented into short fragments (about 200 bp), then the first strand cDNA is synthesized by random hexamer-primer using the mRNA fragments as templates. Buffer, dNTPs, RNase H and DNA polymerase I are added to synthesize the second strand. The double strand cDNA is purified with QiaQuick PCR extraction kit and washed with EB buffer for end repair and poly (A) addition. Finally, sequencing adapters are ligated to the fragments. The fragments are purified by Agarose gel electrophoresis and enriched by PCR amplification. The library products are ready for sequencing analysis via BGISEQ-500.