GSM1923345: miR92a; Drosophila melanogaster; RNA-Seq
Cells were lysed in 1 ml TRI Reagent (Ambion). Following extraction from the lysate, total RNA was cleaned up using the RNeasy Mini kit (Qiagen) and subjected to polyA selection using oligo(dT) Dynabeads (Invitrogen) to isolate mRNA.  Strand-specific mRNA-Seq libraries for Illumina sequencing were prepared as described (Guo et al. 2010), with differences noted below. Briefly, polyA-selected RNA was hydrolyzed in alkaline buffer, resulting in fragments bearing 5'-hydroxyl and 3'-phosphate groups. Fragments between 36-55 nt were size selected and end-specific adapters were sequentially ligated onto each terminus; prior to each ligation step, the appropriate 3' or 5' end chemistry was generated through dephosphorylation or phosphorylation, respectively. Adapter-flanked fragments were reverse transcribed and the resulting cDNA PCR-amplified for 19 cycles using primers complementary to each adapter. The PCR products were purified on a denaturing formamide gel and submitted for deep sequencing.
GEO Accession
GSM1923345