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. , 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 , 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. , Costello Ryan , Damron Weston , Dawood Mariam , DeBuono Nicole , Dimalanta John D. , Edoli Laure , Elango Keerthana , Faustino Nikka , Feng Chao , Ferrari Mathhew , Frankie Keith , Fries Adam , Galloway Anne , Gavrila Vlad , Gemmen Gregory J. , Ghadiali James , Ghorbani Arash , Goddard Logan A. , Guetter Adriana R. , Hendricks Garren L. , Hentschel Jendrik , Honigfort Daniel J. , Hsieh Yun-Ting , Fu Yu-Hsien Hwang , 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 , 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: November 2022

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.