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Improved characterization of single-cell RNA-seq libraries with paired-end avidity sequencing

GSE273222 Homo sapiens Expression profiling by high throughput sequencing 5 samples Submitted 2024/07/30 Platform GPL34295
Summary
Prevailing poly(dT)-primed 3’ single-cell RNA-seq protocols generate barcoded cDNA fragments containing the reverse transcriptase priming site, which is expected to be the poly(A) tail or a genomic adenine homopolymer. Direct sequencing across this priming site was historically difficult because of DNA sequencing errors induced by the homopolymeric primer at the ‘barcode’ end. Here, we evaluate the capability of “avidity base chemistry” DNA sequencing from Element Biosciences to sequence through this homopolymer accurately, and the impact of the additional cDNA sequence on read alignment and precise quantification of polyadenylation site usage. We find that the Element Aviti instrument sequences through the thymine homopolymer into the subsequent cDNA sequence without detectable loss of accuracy.
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Also filed as BioProject PRJNA1140622 and SRA study SRP522501. Searching any of these in the dataset finder brings you back here.

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