GSM9291318: RNA_KO_cPSE_dHPC-F1; Mus musculus; RNA-Seq
Total RNA was extracted using TRIzol-chloroform and the Qiagen RNeasy Mini Kit (74104) with on-column RNase-free DNase (79254) digestion according to manufacturer protocols. Total RNA quality was assessed using the High Sensitivity RNA ScreenTape Assay on an Agilent 4200 TapeStation system. mRNA was isolated from 10 ng total RNA using the NEBNext Poly(A) mRNA Magnetic Isolation Module (E7490) in coordination with NEBNext Multiplex Oligos (E6440) and HighPrep PCR Beads (MagBio AC-60050). Library quality was assessed using the D1000 ScreenTape Assay on an Agilent 4200 TapeStation system. Libraries were quantified using the NEBNext Library Quant Kit (E7630) on an Applied Biosystems QuantStudio 5 Real-Time PCR system. The concentration of amplifiable templates in each library was calculated using the NEBioCalculator qPCR Library Quantification Web Tool. Libraries were then pooled in equal concentrations and submitted for sequencing.