title: IL-17RAfl/fl K13 Cre- Sham replicate 1
design: Total RNA from tngue was used as starting material for deep sequencing         using the Illumina TrueSeq RNA Sample Preparation v2 library preparation kit. Cluster         generation and 75 bp single-end dual-indexed sequencing was performed on Illumina NextSeq         500. Total RNA libraries were generated using Illumina TruSeq Stranded mRNA sample preparation kit. The first step in the workflow involves purifying the poly-A containing mRNA molecules using poly-T oligo attached magneticb eads.Following purification, the mRNA is fragmented into small pieces using divalent cations. The cleaved RNA fragments are copied into first strand cDNA using reverse transcriptase and random primers. Strands pecificity is achieved by using dUTP in the Second Strand Marking Mix, followed by second strand cDNA synthesis using DNA Polymerase I and RNase H. These cDNA fragments then have the addition of a single 'A' base and subsequent ligation of the adapter. The products are then purified and enriched with PCR to create the final cDNA library.  The cDNA libraries are validated using KAPA Biosystems primer premix kit with Illumina-compatible DNA primers and Qubit 2.0 fluorometer. Quality is examined using Agilent Tapestation 2200.The cDNA libraries were pooled at a final concentration 1.8pM. Cluster generation and 75 bp single-read dual-indexed sequencing was performed on Illumina NextSeq 500’s.