GSM5501798: Organoid_day_158_rep2; Homo sapiens; RNA-Seq
Organoids (following 17, 23, 40, 70, 120, 140, or 158 days in culture) were collected for bulk RNA extractions using the RNeasy Mini extraction kit (74104, Qiagen) according to the manufacturer's instructions. RNA quality and concentrations were measured on an Agilent 2100 Bioanalyzer (Agilent). Three replicates were analyzed per time point, every sample containing 1-3 pooled organoids. Sequencing libraries were prepared from a starting amount of 100ng total RNA using the NEBNext® Ultra™ DNA Library Prep Kit for Illumina (E7370L, New England Biolabs) using ribosomal depletion as a selection method. Libraries were sequenced paired-end on an Illumina HiSeq4000 system at the Helmholtz Zentrum Core Facility (Munich, DE). 5 libraries were pooled per lane for a total coverage of on average 50 M reads/library