title: GSM2792487: Patient 1, 24 hour; Homo sapiens; RNA-Seq
library_construction_protocol: Enriched CTCs from the CTC-iChip were spun down, resuspended in RNAlater, flash-frozen in liquid nitrogen, and stored in -80 deg C. RNA was extracted using the Qiagen RNeasy Plus Micro Kit. Using the SMARTer Ultra Low Input RNA Kit for Sequencing – v4 kit (Clontech Laboratories), RNA (~1 ng) was converted to cDNA and amplified per the manufacturer’s protocol. After 18 cycles of second strand synthesis and amplification, cDNA was purified with a 1× Agencourt AMPure XP bead cleanup (Beckman Coulter), and the Nextera® XT DNA Library Preparation kit (Illumina) was used for sample fragmentation/barcoding. Fragmentation used 1 ng of purified cDNA, followed by 12 cycles of amplification prior to unique dual-index barcoding. After the PCR product was purified with a 1.8× Agencourt AMPure XP bead cleanup, the eluted cDNA did not undergo the bead-based normalization step in the Nextera XT protocol.