GSM9439390: bulk_mRNA-seq_MA30_21DPI_B6_brain_3, mRNA-seq; Mus musculus; RNA-Seq
Tissue samples for MA30 and PR8 infection were preserved in 1mL Trizol (15596026; Invitrogen) and extracted with a RNeasy Mini Kit (Cat. No.74104; QIAGEN) according to the manufactures' instructions. Nanodrop was used to test the concentration and purity of the RNA prior to qPCR and bulk RNAseq. Final cDNA libraries containing TruSeq RNA CD indexes (Illumina, 20019792) were quantitated using a Qubit dsDNA HS assay kit (Thermo Fisher Scientific, Q32854). The quality of the libraries was determined by running each on an Agilent TapeStation 4150 using an Agilent D1000 ScreenTape (Agilent: 5067-5582). Smear analysis was performed using Agilent TapeStation Software (Version 4.1.1) with a range of 200-600bp to determine the average size of each library. Size and concentration were then used to calculate the molarity of each library. Libraries were pooled at a final concentration of 750pM with a spike-in of 2% PhiX control library v3 (Illumina, FC-110-3001). A mixture of pooled libraries was loaded on an Illumina NextSeq P1 (300) reagent cartridge (Illumina, 20050264). Paired-end and dual indexing sequence, 150x8x8x150, was performed on NextSeq2000, yielding approximately 20M paired-end reads per sample.
GEO
GSM9439390_r1