title: GSM2640639: NS1_1_118_rep3; Homo sapiens; RNA-Seq
library_construction_protocol: mRNA was prepared using Dynabeads mRNA kit (Invitrogen) and fragmented by heat. For cDNA synthesis, custom oligo-dT primers were used with a barcode and an adapter-linker sequence (CAG ACG TGT GCT CTT CCG ATC T—XXXXXXXX-T15) and with the Affinity Script enzyme (Agilent). After first strand synthesis, samples were pooled together based on ACTB qPCR values. RNA-DNA hybrids was degraded using consecutive acid-alkali treatment. A second sequencing linker (AGA TCG GAA GAG CGT CGT GTAG) was then ligated using T4 ligase (NEB). The mixture was then PCR enriched for 12 cycles and purified to yield final strand specific RNA-seq libraries as previously described55. Libraries were sequenced using a HiSeq 2500 (Illumina) using 50bp x 25bp pair-end sequencing.
attributes.tag: GEO Accession
attributes.value: GSM2640639