Fig. 3: Impact of anchoring methods used to capture oligos to the flow cell on sequence performance.

a Schematic illustration of flow cells. Three different types of binding between capture probes and a flow cell surface were tested as follows; a covalent bond achieved via click chemistry NHS and NH₂ reaction, a non-covalent bond achieved via biotin-avidin but different in the number of biotins per probe, four and one biotin molecules per probe, are shown in the left to the right panels respectively. The same barcode sample shown in Fig. 1 was used for this test. b–d Averaged extension traces for NHS (b), biotin × 4 (c), and biotin × 1 (d), calculated in the same manner shown in Fig. 1f. Colored ribbon plots represent SD at each cycle. All three conditions were repeated twice. e Read length distributions; reads <5 nt were filtered out. f Normalized frequency of stopped reads at each cycle. Note that the value at 24Q corresponds to the fraction of reads that reached 24Q (we were unable to identify whether these reads stopped or extended after this point as 24Q was the last cycle). Colored ribbon plots represent SD. g Cross-correlation of the detected count of 30 types of barcodes between conditions.