GSM1542190: MRS17; Epinephelus coioides; RNA-Seq
Epinephelus coioides larvae (30-days-old)  were homogeneous  whole tissue , RNA was extracted using Trizol reagent, then used RNA zol extracted again to improve quality.  RNeasy Mini Kit (Cat. No. 74106) was used with 1 ug of total RNA for the construction of sequencing libraries. mRNA  is enriched by using the oligo(dT) magnetic beads. Adding the fragmentation buffer, the mRNA is interrupted to 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 sythsize the second strand. The double strand cDNA is purified with QiaQuick PCR extraction kit and washed with EB buffer for end repair and single nucleotide A (adenine) addition. Finally, sequencing adaptors are ligated to the fragments.