alias: GSM4715846
title: GSM4715846: Colon cancer 3N; Homo sapiens; OTHER
library_construction_protocol: Total RNA was extracted using TRIzol (Invitrogen, CA) and RNeasy column methods. LoopSeq sequencing libraries were prepared using LoopSeq Transcriptome Kit according to manufacturer's protocol except when specified. 200 ng of total RNA extracted from cancer patient tissues was reverse transcribed and barcoded using LoopSeq Transcriptome Kit, and an enrichment step using Twist Biosciences probes targeting selected genes annotated to be of significance in cancer development was applied prior to Barcode Amplification in LoopSeq library prep. Specifically, the barcoding reaction was purified using 0.6X SPRISelect ratio and then eluted into 20 μL of Hybridization Mix per manufacturer's pre-capture concentration protocol. The hybridization reactions contain 20 μL of Hybridization Mix, 5 μL of  Blocker Solution, 9 μL of LG Adaptor Blocker (Loop Genomics), 4 μL of Twist Custom Enrichment Panel, 2 μL of Buffer EB (Qiagen), and 30 μL of Hybridization Enhancer. All probe hybridization and washes were conducted per manufacturer's protocol. All target enrichment reagents are from Twist Biosciences except when specified. The captured barcoded cDNA was amplified following the LoopSeq Transcriptome protocol starting from Barcode Amplification.
identifiers.namespace: GEO
identifiers.id: GSM4715846
attributes.tag: GEO Accession
attributes.value: GSM4715846