title: GSM3527362: merged_8; Homo sapiens; RNA-Seq
library_construction_protocol: RNA was extracted using Trizol according to the manufacture's instructions.Specifically, cells were lysed in 1 ml of Trizol, kept at RT for 5 minutes, followed by addition of 200 µl of CHCl3, shaked vigorously for 30", spun @5000 rpm for 2 minutes. The upper phase was recovered and an equal volume of i-PrOH was added, mixed, kept for 5 minutes, and precipitated for 15 minutes @10,000 rpm using Pellet-Paint (Novagen). Library was prepared using Truseq pair-end Illumina kit. Briefly, 10-100 ng of total RNA was reconstituted in water, mixed with Ribosomal removal Buffer containing RNAClean XP beads, and depleted from rRNA. First strand was synthesized using RT, followed by a second strand synthesis, 3' ends were adenylated and ligated to the adaptors, followed by ligateion of the 5' adaptors; the library was purified and measured on DNA-concentration
attributes.tag: GEO Accession
attributes.value: GSM3527362