Journal: bioRxiv
Article Title: Systematic evaluation of 24 extraction and library preparation combinations for metagenomic sequencing of SARS-CoV-2 in saliva
doi: 10.64898/2026.04.16.719115
Figure Lengend Snippet: RNA-Seq libraries were created from the eight different nucleic acid extraction methods using three different RNA-Seq library preparation methods: NEBNext Single Cell/Low Input RNA library preparation kit (NEB), Revelo RNA-Seq High Sensitivity library preparation kit (Revelo), and High Sensitivity BRB-Seq RNA library preparation kit (BRB-Seq). Reads were mapped to viral, bacterial, and eukaryotic genomes present in the RefSeq database. The proportion of total reads mapping to SARS-CoV-2 is shown either grouped by A) nucleic extraction method or B) RNA-Seq library preparation method. As a measure of extraction quality, the proportion of total reads mapping to MS2 is shown either grouped by C) nucleic extraction method or D) by RNA-Seq library preparation method. The proportion of E) SARS-CoV-2 reads or F) MS2 reads to total reads is shown with every combination of nucleic extraction methods and RNA-Seq library preparation kits with a darker shade indicating a higher proportion of SARS-CoV-2 or MS2 reads. (Z1 - Zymo Quick-RNA Magbead, Z2 - Zymo Quick-DNA/RNA Viral Magbead, M1 - MagMAX mirVana Total RNA, M2 - MagMAX Viral/Pathogen, M3 - MagMAX Microbiome Ultra, M4 - MagMAX Viral RNA Isolation, Q1 - QIAamp Viral RNA, P1 - PureLink Viral RNA/DNA). Statistics were calculated by a One-Way ANOVA, followed by Tukey’s HSD post-hoc test. Asterisks above connecting brackets indicate significant differences between those two specific groups. * denotes p<0.05, ** denotes p<0.01, and *** denotes p<0.001.
Article Snippet: The Zymo Quick-DNA/RNA Viral Magbead kit was the only extraction kit that failed to produce detectable RNA via Qubit RNA High Sensitivity Assay Kit in all three positive and negative extracted nucleic acid replicates.
Techniques: RNA Sequencing, Extraction, Single Cell, RNA Library Preparation, Isolation