title: GSM2113396: lipo_II.bam; Homo sapiens; RNA-Seq
library_construction_protocol: RNA was isolated with GeneMATRIX Universal RNA Purification Kit, (cat. no. E3598, EURx), and RNA integrity was assessed using Agilent RNA 6000 Nano Kit on Agilent 2100 Bioanalyzer. All transfections were performed in three independent replicates, numbered I, II and III. Efficacy of PROX1 knockdown was measured with RT-qPCR using specific primers (forward 5'-CCAGCTCCAATATGCTGAAGACCTA-3'; reverse 5'-CATCGTTGATGGCTTGACGTG-3') and Sybr Green chemistry. RNA was subjected to library preparation using Ion AmpliSeq Transcriptome Human Gene Expression Panel, according to manufacturer's protocol. Briefly, RNA was reverse transcribed and cDNA subjected to multiplex PCR reaction to amplify parts of targeted transcripts. Amplicons were then partially digested at primer sequences followed by adapters ligation to amplicons and purification on AMPure® XP beads. Resulting library was  quantified on Bioanalyzer 2100 and diluted to ~100 pM prior to template preparation. Up to eight barcoded libraries were subjected to automated template preparation with Ion PI IC 200 Kit on the Ion Chef Instrument, which performs emulsion PCR on Ion Sphere particles (ISPs), followed by ISPs recovery and template loading on a PI chip. Barcoded RNA-Seq libraries samples were sequenced on a PI chip using the sequencing reagents provided as part of the Ion PI IC 200 Kit, according to the manufacturer’s instructions.
attributes.tag: GEO Accession
attributes.value: GSM2113396