title: GSM8685982: 9N; Mus musculus; RNA-Seq
library_name: GSM8685982
library_construction_protocol: Special pathogen-free (SPF) BALB/c nude mice (n = 15 per group) were maintained in a temperature-controlled facility with 12 h light-dark cycle. BALB/c nude mice (8 wk) were inoculated with mock- and KSHV-infected bone marrow MSCs (1x107 cell count), respectively, by parafemoral injection in the right hindlimb. After 10 weeks, BALB/c nude mice were harvested and subjected to next step experiment. A total amount of 1-3μg RNA per sample was used as input material for the RNA sample preparations. Sequencing libraries were generated using VAHTS Universal V6 RNA-seq Library Prep Kit for Illumina ® （NR604-01/02）following the manufacturer's recommendations and index codes were added to attribute sequences to each sample. Briefly, mRNA was purified from total RNA using poly-T oligo-attached magnetic beads. Then we added fragmentation buffer to break the mRNA into short fragments. First strand cDNA was synthesized using random hexamer primer and RNase H. Second strand cDNA synthesis was subsequently performed using buffer, dNTPs, DNA polymerase I and RNase H. And then, the double stranded cDNA was purified by AMPure P beads or QiaQuick PCR kit. The purified double stranded cDNA was repaired at the end, added a tail and connected to the sequencing connector, then the fragment size was selected, and finally the final cDNA library was obtained by PCR enrichment.