Sequencing by avidity enables high accuracy with low reagent consumption
Authors: Arslan Sinan , Garcia Francisco J. , Guo Minghao , Kellinger Matthew W. , Kruglyak Semyon , LeVieux Jake A. , Mah Adeline H. , Wang Haosen , Zhao Junhua , Zhou Chunhong , Altomare Andrew , Bailey John , Byrne Matthew B. , Chang Chiting , Chen Steve X. , Cho Byungrae , Dennler Claudia N. , Dien Vivian T. , Fuller Derek , Kelley Ryan , Khandan Omid , Klein Michael G. , Kim Michael , Lajoie Bryan R. , Lin Bill , Liu Yu , Lopez Tyler , Mains Peter T. , Price Andrew D. , Robertson Samantha R. , Taylor-Weiner Hermes , Tippana Ramreddy , Tomaney Austin B. , Zhang Su , Abtahi Minna , Ambroso Mark R. , Bajari Rosita , Bellizzi Ava M. , Benitez Chris B. , Berard Daniel R. , Berti Lorenzo , Blease Kelly N. , Blum Angela P. , Boddicker Andrew M. , Bondar Leo , Brown Chris , Bui Chris A. , Calleja-Aguirre Juan , Cappa Kevin , Chan Joshua , Chang Victor W. , Charov Katherine , Chen Xiyi , Constandse Rodger M. , Damron Weston , Dawood Mariam , DeBuono Nicole , Dimalanta John D. , Edoli Laure , Elango Keerthana , Faustino Nikka , Feng Chao , Ferrari Matthew , Frankie Keith , Fries Adam , Galloway Anne , Gavrila Vlad , Gemmen Gregory J. , Ghadiali James , Ghorbani Arash , Goddard Logan A. , Guetter Adriana Roginski , Hendricks Garren L. , Hentschel Jendrik , Honigfort Daniel J. , Hsieh Yun-Ting , Hwang Fu Yu-Hsien , Im Scott K. , Jin Chaoyi , Kabu Shradha , Kincade Daniel E. , Levy Shawn , Li Yu , Liang Vincent K. , Light William H. , Lipsher Jonathan B. , Liu Tsung-li , Long Grace , Ma Rui , Mailloux John M. , Mandla Kyle A. , Martinez Anyssa R. , Mass Max , McKean Daniel T. , Meron Michael , Miller Edmund A. , Moh Celyne S. , Moore Rachel K. , Moreno Juan , Neysmith Jordan M. , Niman Cassandra S. , Nunez Jesus M. , Ojeda Micah T. , Ortiz Sara Espinosa , Owens Jenna , Piland Geoffrey , Proctor Daniel J. , Purba Josua B. , Ray Michael , Rong Daisong , Saade Virginia M. , Saha Sanchari , Tomas Gustav Santo , Scheidler Nicholas , Sirajudeen Luqmanal H. , Snow Samantha , Stengel Gudrun , Stinson Ryan , Stone Michael J. , Sundseth Keoni J. , Thai Eileen , Thompson Connor J. , Tjioe Marco , Trejo Christy L. , Trieger Greg , Truong Diane Ni , Tse Ben , Voiles Benjamin , Vuong Henry , Wong Jennifer C. , Wu Chiung-Ting , Yu Hua , Yu Yingxian , Yu Ming , Zhang Xi , Zhao Da , Zheng Genhua , He Molly , Previte Michael 
Published: May 2023

We present avidity sequencing, a sequencing chemistry that separately optimizes the processes of stepping along a DNA template and that 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. Avidity sequencing achieves high accuracy, with 96.2% and 85.4% of base calls having an average of one error per 1,000 and 10,000 base pairs, respectively. We show that the average error rate of avidity sequencing remained stable following a long homopolymer.