GSM9111285: PD_SP_R1; Mus musculus; RNA-Seq
0.5x106 cells were collected on day 5 of EB culture and mesodermal populations were FACS sorted using FLK and PDGFRa surface expression. Subsequently RNA was isolated using Qiagen RNAeasy kit (Qiagen cat. 74134). The quality of the isolated RNA was determined with a Fragment Analyzer (Agilent, Santa Clara, California, USA). The TruSeq Stranded mRNA protocol (Illumina, Inc, California, USA) was used. Briefly, total RNA samples (100-1000 ng) were poly A enriched and then reverse-transcribed into double-stranded cDNA. The cDNA samples were fragmented, end-repaired and adenylated before ligation of TruSeq adapters containing unique dual indices (UDI) for multiplexing. Fragments containing TruSeq adapters on both ends were selectively enriched with PCR. The quality and quantity of the enriched libraries were validated using the Fragment Analyzer. The libraries were normalized to 10 nM in Tris-Cl 10 mM, pH8.5 with 0.1% Tween 20.