
This SARS-CoV-2 targeted, a rapid viral RNA library preparation kit compatible with Illumina sequencers, provides an approximately five hour turn-around time from viral RNA sample to sequencing. The kit includes cDNA synthesis, SARS-CoV-2 enrichment, and library amplification and indexing reagents. The Enhanced protocol removes the need for normalization and quantification prior to amplification and indexing. RNA input is 5 μl, regardless of SC2 viral load in the sample extract. Amplification is performed using two primer pools that produce overlapping amplicons that are approximately 250 bp in size; as a result, no fragmentation is necessary prior to sequencing on Illumina platforms. 10 bp unique dual indexes (UDIs) are provided in the kit, and with a cycle read length of 149 for each paired-end read, the sequencing time is just over 28 hours. Version Updates: V2- Recommending 1:10 dilution of index primer on low viral titer samples to reduce short fragments. Controlled vocabulary for the use of positive control and negative template control has been added to the Sample Sheet following miseq loading guidelines.
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