QIAseq DIRECT SARS-CoV-2 Kit B

A targeted next-generation sequencing library preparation kit from QIAGEN for SARS-CoV-2 research. The QIAseq DIRECT SARS-CoV-2 Kit B uses primer pool B to amplify defined regions of the viral genome in a direct library preparation workflow compatible with Illumina sequencing platforms. The reagents support reproducible, multiplexed sample processing and are intended for research use, complementing other primer pools to broaden genome coverage.

An enhanced protocol published by FDA CFSAN GenomeTrakr researchers describes a workflow providing an approximately five hour turnaround time from viral RNA sample to sequencing. The kit includes cDNA synthesis, SARS-CoV-2 enrichment, and library amplification and indexing reagents, with an RNA input of 5 μl regardless of viral load. Amplification uses two primer pools that produce overlapping amplicons approximately 250 bp in size, eliminating the need for fragmentation prior to sequencing on Illumina platforms. The kit provides 10 bp unique dual indexes, and with a cycle read length of 149 for each paired-end read, sequencing time is just over 28 hours.

Components supplied include RP Primer, EZ Reverse Transcriptase, RNase Inhibitor, Multimodal RT Buffer (5×), QIAseq 2X HiFi MM, DIRECT SARS-CoV-2 Pool 1, DIRECT SARS-CoV-2 Pool 2, a QIAseq DIRECT UDI Set, Nuclease-Free Water, and QIAseq Beads.