design: Tumor total RNA were purified from the same sample using the AllPrep DNA/RNA/miRNA kit (QIAGEN) with disruption on a Tissuelyser II (QIAGEN). RNA integrity was verified on an Agilent 2100 Bioanalyzer using RNA Nano reagents (Agilent Technologies).Transcriptome libraries were prepared using 200.1,000 ng of total RNA. PolyA+ RNA isolation, cDNA synthesis, end-repair, A-base addition, and ligation of the Illumina indexed adapters were performed according to the TruSeq RNA protocol (Illumina). Libraries were size selected for 250.300 bp cDNA fragments on a 3%Nusieve 3:1 (Lonza) gel, recovered using QIAEX II reagents (QIAGEN), and PCR amplified using Phusion DNA polymerase (New England Biolabs). Library quality was measured on an Agilent 2100 Bioanalyzer for product size and concentration.
identifiers.namespace: UM-MCTP
identifiers.id: UT_4059_MCTP_SI_17737