Sequencing by avidity enables high accuracy with low reagent consumption

Arslan, S.; Garcia, F. J.; Guo, M.; Kellinger, M. W.; Kruglyak, S.; LeVieux, J. A.; Mah, A. H.; Wang, H.; Zhao, J.; Zhou, C.; Altomare, A.; Bailey, J.; Byrne, M. B.; Chang, C.; Chen, S. X.; Dennler, C. N.; Dien, V. T.; Fuller, D.; Kelley, R.; Khandan, O.; Klein, M. G.; Kim, M.; Lajoie, B. R.; Lin, B.; Liu, Y.; Lopez, T.; Mains, P. T.; Price, A. D.; Robertson, S. R.; Taylor-Weiner, H.; Tippana, R.; Tomaney, A. B.; Zhang, S.; Ambroso, M. R.; Bajari, R.; Bellizzi, A. M.; Benitez, C. B.; Berard, D. R.; Berti, L.; Blease, K. N.; Blum, A. P.; Boddicker, A. M.; Bondar, L.; Brown, C.; Bui, C. A.; Call

doi:10.1101/2022.11.03.514117

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bioRxiv · 10.1101/2022.11.03.514117

Sequencing by avidity enables high accuracy with low reagent consumption

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Abstract

We present avidity sequencing - a novel sequencing chemistry that separately optimizes the process of stepping along a DNA template and the process of identifying each nucleotide within the template. Nucleotide identification uses multivalent nucleotide ligands on dye-labeled cores to form polymerase-polymer nucleotide complexes bound to clonal copies of DNA targets. These polymer-nucleotide substrates, termed avidites, decrease the required concentration of reporting nucleotides from micromolar to nanomolar, and yield negligible dissociation rates. We demonstrate the use of avidites as a key component of a sequencing technology that surpasses Q40 accuracy and enables a diversity of applications that include single cell RNA-seq and whole human genome sequencing. We also show the advantages of this technology in sequencing through long homopolymers.

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Arslan, S., Garcia, F. J., Guo, M., Kellinger, M. W., Kruglyak, S., LeVieux, J. A., Mah, A. H., Wang, H., Zhao, J., Zhou, C., Altomare, A., Bailey, J., Byrne, M. B., Chang, C., Chen, S. X., Dennler, C. N., Dien, V. T., Fuller, D., Kelley, R., Khandan, O., Klein, M. G., Kim, M., Lajoie, B. R., Lin, B., Liu, Y., Lopez, T., Mains, P. T., Price, A. D., Robertson, S. R., Taylor-Weiner, H., Tippana, R., Tomaney, A. B., Zhang, S., Ambroso, M. R., Bajari, R., Bellizzi, A. M., Benitez, C. B., Berard, D. R., Berti, L., Blease, K. N., Blum, A. P., Boddicker, A. M., Bondar, L., Brown, C., Bui, C. A., Call. 2022-11-04. Sequencing by avidity enables high accuracy with low reagent consumption. https://doi.org/10.1101/2022.11.03.514117

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