DMSO treatment with influenza
Total RNA (5 to 10 g) with a Bioanalyzer RIN score of greater than 8.0 was used for library preparation. Removal of ribosomal RNA was done by poly-A selection utilizing Oligo-dT beads (mRNA Direct kit, Life Technologies). Fragmentation of mRNA was done in reverse transcriptase buffer by heating to 94C for 8 minutes. cDNA preparation was done using random hexamers and SuperScript III RT enzyme (Life Technologies, per manufacturers instructions). DS-cDNA was prepared by a second-strand reaction. The cDNA thus prepared was blunt ended and had 3 ends with A base added. The Illumina adapters were then ligated to the ends. Unique dual index tags were introduced into the ligated fragments by specific primers (12-15 cycle amplification).  The amplified fragments were then sequenced on an Illumina NovaSeq-6000 using paired end reads extending 150 bases