GSM2649461: Prp31_P18_rep2; Drosophila melanogaster; RNA-Seq
Total RNA was extracted using the RNAeasy Mini kit (Qiagen) according to the manufacturer's instructions. mRNA was isolated from 1ug total RNA by poly-dT enrichment using the   NEBNext Poly(A) mRNA Magnetic Isolation Module according to the   manufacturers instructions. Final elution was done in 15ul 2x first   strand cDNA synthesis buffer (NEBnext, NEB). After chemical   fragmentation by incubating for 15 min at 94°C the sample was directly   subjected to the workflow for strand specific RNA-Seq library   preparation (Ultra Directional RNA Library Prep, NEB). For ligation   custom adaptors were used (Adaptor-Oligo 1:   5'-ACA-CTC-TTT-CCC-TAC-ACG-ACG-CTC-TTC-CGA-TCT-3', Adaptor-Oligo 2:   5'-P-GAT-CGG-AAG-AGC-ACA-CGT-CTG-AAC-TCC-AGT-CAC-3'). After ligaton   adapters were depleted by an XP bead purification (Beckman Coulter)   adding beads in a ratio of 1:1. Indexing was done during the following   PCR enrichment (15 cycles) using custom amplification primers carring   the index sequence indicated with ‘NNNNNN’. (Primer1: Oligo_Seq    AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCT, primer2:   GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT, primer3: CAAGCAGAAGACGGCATACGAGAT   NNNNNN GTGACTGGAGTT. After two more XP beads purifications (1:1)   libraries were quantified using Qubit dsDNA HS Assay Kit (Invitrogen).   For Illumina flowcell production, samples were equimolarly pooled and   distributed on one lane for 75bp single end sequencing on Illumina HiSeq   2500 resulting in on average 28 Mio reads per sample.
GEO Accession
GSM2649461