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. 2022 Aug 13;14(1):2106621. doi: 10.1080/19420862.2022.2106621

Figure 4.

(a) Electrophoresis showing bands of DNA ranging in size from 600 to 3500 kilobases for cDNA libraries I, II, III, and IV. (b Bar graph showing >500,000 pass reads for cDNA libraries I, II, III, and IV, and ~300,000 un/misclassified reads. (c) Graph showing that average sequence read length is >1000 for cDNA libraries I and II and <1000 for cDNA libraries III and IV. (d) Table of sequence characteristics for the heavy- and light-chain sequences obtained from cDNA libraries I, II, III, and IV.

NAb-seq of four single B cells revealed antibody sequences. (a) cDNA library size and amplification. Total RNA from four sorted single B cells were extracted and converted to whole transcriptome cDNA libraries I–IV. (b) Basecalling in Guppy’s super-high accuracy mode yielded approximately 5 million total pass reads. (c) Read length of pass reads varied from ~400 bp to ~5000 bp. (d) Sequence analysis of cDNA libraries I to IV reveals V(D)J recombination, C gene usage and complementarity-determining region 3 (CDR3) amino acid sequence. Consensus calling of antibody transcripts revealed IgM and IgD isotypes.