GSM6482209: Normal SAP development D23, Bio_Rep4 , Bulk- RNAseq; Homo sapiens; RNA-Seq
Cell pellets were snapfrozen for analysis at selected time-points over the differentiation process. To extract RNA from the cells, phase separation in cells lysed in TRIzol Reagent was induced with chloroform. RNA was precipitated with isopropanol and linear acrylamide and washed with 75% ethanol. The samples were resuspended in RNase-free water. After RiboGreen quantification and quality control by Agilent BioAnalyzer, 125-500ng of total RNA with an RNA integrity number varying from 7.8-10 underwent polyA selection and TruSeq library preparation according to instructions provided by Illumina, with 8 cycles of PCR. Samples were barcoded and run on a HiSeq 4000 in a 50bp/50bp paired end run, using the HiSeq 3000/4000 SBS Kit.