GSM3036929: RNA-seq 5 hpO OMIS rep2; Danio rerio; RNA-Seq
Total RNA of oocytes were purified using RNeasy Mini kit (QIAGEN, 74104) as recommended protocols First stand cDNA was synthesized using a commercial kit (Invitrogen, Cat. 18080-051). Second strand cDNA was synthesized with second strand synthesis buffer (Invitrogen, Cat. 10812014), MgCl2, DTT, dNTP, dUTP, RNase H (Fermentas, Cat. EN0202), E. coli DNA ligase (NEB, Cat. M0205S) and DNA polymerase I (NEB, Cat. M0209S). DNA was purified after 2-hour incubation on thermomixer at 16°C. Synthesized cDNA was subjected to library preparation. DNA was end-repaired, adenylated, and ligated to TruSeq sequencing adaptors. Libraries were then treated with UDG (AMPErase, Cat. N808-0096) at 37°C for 1 hour followed by DNA amplification using Phusion HF DNA polymerase (NEB, Cat. M0530L). The amplified DNA was size selected using AMPure Beads for 200-500bp DNA fragments.
GEO Accession
GSM3036929