Pooled RNA extraction and PCR assay for efficient SARS-CoV-2 detection

Roni Ben-Ami; Agnes Klochendler; Matan Seidel; Tal Sido; Ori Gurel-Gurevich; Moran Yassour; Eran Meshorer; Gil Benedek; Irit Fogel; Esther Oiknine-Djian; Asaf Gertler; Zeev Rotstein; Bruno Lavi; Yuval Dor; Dana G Wolf; Maayan Salton; Yotam Drier

For matrix pool design we pooled equal volumes of sample lysate from each of the subjects to a final volume of 450 µL and used MagNA Pure 96 kit (Roche Lifesciences) using Roche platform. As supply was limited for this kit, we have used QIAsymphony DSP Virus/Pathogen kit on Qiasymphony platform for 1:8 pool design. We pooled equal volumes of sample lysate from each of the subjects to a final volume of 400 µL. Positive 1:8 pools were validated by single tests using QIAsymphony RNA kit on Qiasymphony platform. Both Qiagen kits were used with Zymo lysis buffer, and therefore we skipped the lysis and Proteinase K step. RNA was eluted into 60 µL; 10 µL of RNA was used for a 30 µL reaction using Real-Time Fluorescent RT-PCR kit (BGI). . CC-BY-NC 4.0 International license It is made available under a author/funder, who has granted medRxiv a license to display the preprint in perpetuity.