RNA-Seq of Brugia pahangi recovered from vehicle treated jirds 1 week post-first dose: pooled adult females
cDNA libraries were produced from the low-input RNA using the Takara SMART-Seq v4 Ultra-low input RNA kit. An S220 Focused-Ultrasonicator (Covaris) was used to fragment the DNA, and library preparation was performed using the KAPA hyper prep kit for DNA (KK8504). Truncated universal stub adapters were used for ligation, and indexed primers were used during PCR amplification to complete the adapters and to enrich the libraries for adapter-ligated fragments. Samples were checked for quality on an AATI (now Agilent) Fragment Analyzer. Samples were then transferred to the Vincent J. Coates Genomics Sequencing Laboratory (GSL), another QB3-Berkeley Core Research Facility at UC Berkeley, where Illumina sequencing libraries were quantitative PCR was used to calculate sequence-able molarity with the Kapa Biosystems Illumina Quant qPCR Kits on a BioRad CFX Connect thermal cycler. Libraries were then pooled evenly by molarity and sequenced on an Illumina NovaSeq6000 150PE S4 flowcell, generating 25M read pairs per sample.