Deep amplicon sequencing of mus musculus: male cerebellum targeting virus
Four weeks post injection, brains were dissected in 5 mM HEPES in HBSS and total RNA was isolated from individual brain regions using QIAzol reagent (Qiagen, #79306) and the Qiagen miRNeasy Kit reagent (Qiagen, #217004) according to the manufacturers instructions. Purified RNA was delivered to the Massively Parallel Sequencing Shared Resource Core (MPSSR) at the Oregon Health and Science University for Illumina library preparation and then transferred to the Molecular Technologies Core (MTC) at the Oregon National Primate Research Center for Illumina AmpliconSequencing. The library was prepared with the TruSeq DNA Nano Library Prep Kit and sequenced with a MiSeq instrument (1 M reads, 500 cycles, 2x500bp pairedend; Illumina). 2nM pooled libraries were spiked with 5% PhiX prior to the sequencing run.