GSM7062065: biol WT2; Mus musculus; RNA-Seq
To isolate RNA, mouse cortex was homogenized in 1mL of TRIzol and RNA was purified using chloroform and Isopropyl alcohol Poly(A)+ transcripts were isolated with the NEBNext Poly(A) mRNA Magnetic Isolation Module kit according to the manufacturer's protocol (New England BioLabs Inc., Massachusetts, USA). RNA samples were randomly primed (5' d(N6) 3' [N=A,C,G,T]) and fragmented. The first strand was synthesized with Protoscript II Reverse Transcription with a modified extension period as to what is stated in the manufacturer's protocol (30 minutes at 42⁰C). The remaining steps of library generation were carried out according to the manufacturer's protocol NEBNext® Ultra™ II Directional RNA Library Prep Kit for Illumina® (New England BioLabs Inc., Massachusetts, USA).