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Isoform-Level Analysis of 10x Genomics Single-Cell cDNA Libraries from Cultured K562 Cells Using Long-Read Sequencing

Bachmann, J.; Olsen, R.-A.; Bhatt, A.; Crang, N.; Contreras-Lopez, O.; Mezger, A.

2025-08-15 genomics
10.1101/2025.08.12.668929 bioRxiv
Show abstract

Integration of Oxford Nanopore Technologies (ONT) long-read sequencing with 10x Genomics single-cell cDNA libraries enables novel transcript detection, isoform analysis and captures full-length gene body coverage. The purpose of the study was the comparison of three approaches for sequencing 10x Genomics Chromium Single Cell cDNA libraries using long-read sequencing: single-cell full-length transcript sequencing by sampling (FLT-seq), the cDNA-PCR Sequencing Kit (SQK-PCS111) and the PCR Expansion Kit (EXP-PCA001). Our aim was to evaluate their efficiency in enriching full-length cDNA fragments, identifying barcodes, detecting novel isoforms and mutations, and characterizing transcript coverage profiles.

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