The 25 samples that underwent RNA-sequencing had an average of 63 million paired-reads per sample, and were within the expected purity ratio of ≥1.8. The mean (SD) RNA yield was 772.3 (939.4) ng., and the mean RNA integrity number for all 25 samples was ≥ 6.0. On average 93% of reads had a quality score of 30 or higher. Samples were mapped to the hg19 Human genome build with an average of 97% mapping rate.


Syracuse University

"RNA extraction was performed using the RNeasy Plus Mini Kit (Qiagen) following the manufacturer’s protocol from sample prepared from the tip of the swab. RNA purity was assessed using Qubit 2.0 fluorometer and Agilent TapeStation 2200 for RNA quantity and quality. Host RNA-seq libraries were generated using Illumina TruSeq RNA Access, which converts total RNA into template molecules, followed by sequence-specific capture of coding RNA. The complementary DNA (cDNA) libraries were quantified using Qubit 2.0 fluorometer, and the quality was examined using Agilent Tapestation 2200.The cDNA libraries were pooled at a final concentration of 2.0 pM. Cluster generation and paired-read, dual-indexed 75 bp sequencing was performed on Illumina NextSeq 500."

Method for RNA Sequencing with NP Swab

Lopez, S.M.C., Martin, J.M., Johnson, M. et al. A method of processing nasopharyngeal swabs to enable multiple testing. Pediatr Res 86, 651–654 (2019). https://doi-org.libezproxy2.syr.edu/10.1038/s41390-019-0498-1
https://www.nature.com/articles/s41390-019-0498-1

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