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Fig. 4 | Genome Medicine

Fig. 4

From: BALDR: a computational pipeline for paired heavy and light chain immunoglobulin reconstruction in single-cell RNA-seq data

Fig. 4

Reconstruction of Ig transcripts by BALDR is highly accurate. The fidelity of bioinformatic reconstruction of immunoglobulin variable regions was assessed by sequence comparison to a “gold-standard” sequence obtained independently from an aliquot of the single B cell lysate prior to amplification. a Accuracy, defined as correct identification of clonotype (V(D)J gene segment and CDR3 sequence of NGS-reconstructed IgH and IgL relative to 115 IgH and 140 IgL sequences obtained from nested RT-PCR and Sanger sequencing for all filtering methods. b Clonal distribution of single cells. The cells were assigned into families based on V, J, and CDR3 length of IgH and IgL. c Assessment of NGS-reconstruction fidelity at the nt level. Nucleotide sequences of reconstructed IgH chains determined to be accurate at the clonotype level were compared to matched sequences obtained by Sanger sequencing by blastn alignment. d SHMs in V region compared to germline IMGT sequences

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