Repertoire Sequencing / DNA Sequencing of Immunoglobulin Genes / Molecular Amplification fingerprinting

Rep-Seq is a collective term for repertoire sequencing technologies (Benichou et al., 2012), including Ig-seq (Georgiou et al., 2014) and MAF (Khan et al., 2016). Ig-seq is a targeted gDNA amplification method performed with primers complementary to the rearranged V-region gene (VDJ recombinant). Amplification of gDNA is then performed with the appropriate 5ê primers. Although throughput is high, information regarding which VH and VL chains were paired in the same cell is lost, as cells are lysed in bulk and VH and VL genes are amplified in separate reactions. MAF (illustrated below) was developed to correct the amplification biases in Ig-seq, by using tagging with unique identifiers (UIDs) before and during the multiplex PCR amplification, as well as a bioinformatics pipeline.


  • Measures antibody frequencies with up to 99% accuracy



Illumina Library prep and Array Kit Selector


Hackl H., Charoentong P., Finotello F. and Trajanoski Z. Computational genomics tools for dissecting tumour-immune cell interactions. Nat Rev Genet. 2016;17:441-458

Hou X. L., Wang L., Ding Y. L., Xie Q. and Diao H. Y. Current status and recent advances of next generation sequencing techniques in immunological repertoire. Genes Immun. 2016;17:153-164

Georgiou G., Ippolito G. C., Beausang J., Busse C. E., Wardemann H. and Quake S. R. The promise and challenge of high-throughput sequencing of the antibody repertoire. Nat Biotechnol. 2014;32:158-168


Khan T. A., Friedensohn S., Gorter de Vries A. R., Straszewski J., Ruscheweyh H. J. and Reddy S. T. Accurate and predictive antibody repertoire profiling by molecular amplification fingerprinting. Sci Adv. 2016;2:e1501371