title: GSM4291728: PDC_Day8.1; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA samples for RNA sequencing were extracted from PHHs cultured in the μHEP slides and a variety of commercial microtiter plates (Corning® BiocoatTM collagen-coated 384-well plate, Cat No: Cat No: 356664, Corning; Falcon® 24-well Clear Flat Bottom Tissue Culture-treated Multiwell Cell Culture Plate, Cat No: 353047, Corning; and a 12 Well glass bottom plate with high performance #1.5 cover glass, Cat No: P12-1.5H-N, In Vitro Scientific, CA, USA). Trizol was added to each sample, incubated for 5 minutes and stored at -80 °C until further isolation. Total RNA was isolated using TRIzol reagent (Thermo Fisher Scientific) following the manufacturer's instructions using chloroform, isopropanol, and 75% ethanol. The resulting RNA pellet was resuspended in DEPCtreated water. RNA quantification was performed by Qubit (Thermo Fisher Scientific), RIN values were determined using a TapeStation 2200 (Agilent Technologies, Santa Clara, CA) and A260/280 and A260/230 ratios were assessed using the NanoDrop® Spectrophotometer ND-1000 (Thermo Fisher Scientific). For each sample, 0.5-1.0 μg total RNA with a RIN value greater than 7.0, A260/280 ratio greater than 1.8, and A260/230 ratio greater than 2.0 was used for library preparation. 0.5 μg - 1.0 μg total RNA of each sample was used for library preparation. Libraries were prepared by using KAPA Stranded RNA-Seq Kit. Library quantification was conducted by qPCR and TapeStation (Agilent Technologies) measurements. Libraries were subsequently pooled and sequenced on Illumina HiSeq 4000 with a 150bp pair-end run.
attributes.tag: GEO Accession
attributes.value: GSM4291728