title: GSM3753338: H293T_ILF3OE_Rep1T2; Homo sapiens; OTHER
library_construction_protocol: RNA was extracted Total RNA was extracted from cells using Qiagen Micro or Mini RNeasy kit (Qiagen cat:74004 or 74104).using RNAdvanced Tissue Kit (Beckman Coulter) followed by TURBO DNase treatment (Thermo Fisher). The mmPCR–seq was performed as described in (Zhang et al 2014 ). Briefly, total RNA is extracted from cells using a Qiagen Micro or Mini RNeasy kit (Qiagen cat:74004 or 74104) and reverse transcribed using iScript Advanced reverse transcriptase (Bio-Rad). The cDNAs were purified using Ampure XP Beads (Beckman Coulter), with an elution volume of 10 ul. 250 ng of cDNA was used in a preamplification reaction and amplified cDNA was purified using Ampure XP beads and eluted in 10 ul.  50 ng of pre-amplified cDNA was loaded into each well of an Access Array microfluidic chip (Fluidigm). The PCR reactions were performed on the Access Array System (Fluidigm) using KAPA2G 5X Fast Multiplex PCR Mix (Kapa Biosystems). Barcodes were added in a second round of PCR using Phusion DNA polymerase (NEB cat: M0531S). Samples were sequenced with 76 base-pair paired-end reads using an Illumina NextSeq (Illumina, San Diego, CA). mmPCR-seq