title: GSM7675183: skin explant day3 ZIKV.buffer_1; Homo sapiens; RNA-Seq
library_construction_protocol: The RNA quality check, sequencing, and bioinformatic analysis were performed at Azenta Life Sciences (South Plainfield). RNA samples were quantified using Qubit 2.0 Fluorometer (Life Technologies, Carlsbad, CA, USA) and RNA integrity was checked using Agilent TapeStation 4200 (Agilent Technologies, Palo Alto, CA, USA). RNA sequencing libraries were prepared using the NEBNext Ultra II RNA Library Prep Kit for Illumina using the manufacturer's instructions (NEB, Ipswich, MA, USA). Briefly, mRNAs were first enriched with Oligo(dT) beads.  Enriched mRNAs were fragmented for 15 minutes at 94 °C. First-strand and second-strand cDNAs were subsequently synthesized. cDNA fragments were end-repaired and adenylated at 3'ends, and universal adapters were ligated to cDNA fragments, followed by index addition and library enrichment by limited-cycle PCR. The sequencing libraries were validated on the Agilent TapeStation (Agilent Technologies, Palo Alto, CA, USA), and quantified by using Qubit 2.0 Fluorometer (Invitrogen, Carlsbad, CA) as well as by quantitative PCR (KAPA Biosystems, Wilmington, MA, USA). The sequencing libraries were clustered on flow cells. After clustering, the flow cells were loaded onto the Illumina HiSeq instrument (4000 or equivalent) according to the manufacturer's instructions.  The samples were sequenced using a 2x150bp Paired-End (PE) configuration. Image analysis and base calling were conducted by the HiSeq Control Software (HCS).