RNA-Seq (polyA+) analysis of ALL sample TARGET-10-PAKKCA-03A-01R
PolyA+ RNA was purified using the MACS mRNA isolation kit (Miltenyi Biotec, Bergisch Gladbach, Germany), from 5-10ug of DNaseI-treated total RNA as per the manufacturer’s instructions. Double-stranded cDNA was synthesized from the purified polyA+ RNA using the Superscript Double-Stranded cDNA Synthesis kit (Invitrogen, Carlsbad, CA, USA) and random hexamer primers (Invitrogen) at a concentration of 5µM. The cDNA was fragmented by sonication and a paired-end sequencing library prepared following the Illumina paired-end library preparation protocol (Illumina, Hayward, CA, USA).