title: GSM3730699: G155-24; Homo sapiens; RNA-Seq
library_construction_protocol: Surgical specimens were transferred immediately from the operating room to the surgical pathology lab where a section of full-thickness sigmoid colon was obtained from a site of active inflammation and stored in RNAlater. RNA was extracted from tissue samples using trizol reagent followed by purification with the Qiagen RNeasy Mini Kit. The cDNA libraries were prepared using the NEXTflex™ Illumina Rapid Directional RNA-Seq Library Prep Kit (BioO Scientific) as per the manufacturer's instructions. Briefly, polyA RNA was purified from 200 ng of total RNA using oligo (dT) beads. The extracted mRNA fraction was subjected to fragmentation, reverse transcription, end repair, 3'– end adenylation, and adaptor ligation, followed by PCR amplification and SPRI bead purification (Beckman Coulter). The unique index sequences were incorporated in the adaptors for multiplexed high-throughput sequencing. The final product was assessed for its size distribution and concentration using BioAnalyzer High Sensitivity DNA Kit (Agilent Technologies). Pooled libraries were diluted to 2 nM in EB buffer (Qiagen) and then denatured using the Illumina protocol. The denatured libraries were diluted to 10 pM by pre-chilled hybridization buffer and loaded onto a TruSeq v2 Rapid flow cell on an Illumina HiSeq 2500 and run for 50 cycles using a single-read recipe according to the manufacturer's instructions.
attributes.tag: GEO Accession
attributes.value: GSM3730699