Sequencing by avidity enables high accuracy with low reagent consumption.
Sinan ArslanFrancisco J GarciaMinghao GuoMatthew W KellingerSemyon KruglyakJake A LeVieuxAdeline H MahHaosen WangJunhua ZhaoChunhong ZhouAndrew AltomareJohn BaileyMatthew B ByrneChiting ChangSteve X ChenByungrae ChoClaudia N DennlerVivian T DienDerek FullerRyan KelleyOmid KhandanMichael G KleinMichael KimBryan R LajoieBill LinYu LiuTyler LopezPeter T MainsAndrew D PriceSamantha R RobertsonHermes Taylor-WeinerRamreddy TippanaAustin B TomaneySu ZhangMinna AbtahiMark R AmbrosoRosita BajariAva M BellizziChris B BenitezDaniel R BerardLorenzo BertiKelly N BleaseAngela P BlumAndrew M BoddickerLeo BondarChris BrownChris A BuiJuan Calleja-AguirreKevin CappaJoshua ChanVictor W ChangKatherine CharovXiyi ChenRodger M ConstandseWeston DamronMariam DawoodNicole DeBuonoJohn D DimalantaLaure EdoliKeerthana ElangoNikka FaustinoChao FengMatthew FerrariKeith FrankieAdam FriesAnne GallowayVlad GavrilaGregory J GemmenJames GhadialiArash GhorbaniLogan A GoddardAdriana Roginski GuetterGarren L HendricksJendrik HentschelDaniel J HonigfortYun-Ting HsiehYu-Hsien Hwang FuScott K ImChaoyi JinShradha KabuDaniel E KincadeShawn LevyYu LiVincent K LiangWilliam H LightJonathan B LipsherTsung-Li LiuGrace LongRui MaJohn M MaillouxKyle A MandlaAnyssa R MartinezMax MassDaniel T McKeanMichael MeronEdmund A MillerCelyne S MohRachel K MooreJuan MorenoJordan M NeysmithCassandra S NimanJesus M NunezMicah T OjedaSara Espinosa OrtizJenna OwensGeoffrey PilandDaniel J ProctorJosua B PurbaMichael RayDaisong RongVirginia M SaadeSanchari SahaGustav Santo TomasNicholas ScheidlerLuqmanal H SirajudeenSamantha SnowGudrun StengelRyan StinsonMichael J StoneKeoni J SundsethEileen ThaiConnor J ThompsonMarco TjioeChristy L TrejoGreg TriegerDiane Ni TruongBen TseBenjamin VoilesHenry VuongJennifer C WongChiung-Ting WuHua YuYingxian YuMing YuXi ZhangDa ZhaoGenhua ZhengMolly HeMichael PrevitePublished in: Nature biotechnology (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.