High-quality full-length immunoglobulin profiling with unique molecular barcoding

Varování

Publikace nespadá pod Ekonomicko-správní fakultu, ale pod Středoevropský technologický institut. Oficiální stránka publikace je na webu muni.cz.
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TURCHANINOVA M. A. DAVYDOV Alexey Nikolayevich BRITANOVA O. V. SHUGAY Mikhail BIKOS Vasileios EGOROV Evgeny KIRGIZOVA V. I. MERZLYAK E. M. STAROVEROV D. B. BOLOTIN D. A. MAMEDOV Ilgar IZRAELSON Mark LOGACHEVA M. D. KLADOVA O. PLEVOVÁ Karla POSPÍŠILOVÁ Šárka CHUDAKOV Dmitriy

Rok publikování 2016
Druh Článek v odborném periodiku
Časopis / Zdroj NATURE PROTOCOLS
Fakulta / Pracoviště MU

Středoevropský technologický institut

Citace
www http://www.nature.com/nprot/journal/v11/n9/full/nprot.2016.093.html#access
Doi http://dx.doi.org/10.1038/nprot.2016.093
Obor Biochemie
Klíčová slova ZEBRAFISH ANTIBODY REPERTOIRE; B-CELL REPERTOIRE; RHEUMATOID-ARTHRITIS; IMMUNE REPERTOIRES; MULTIPLEX PCR; MEMORY; VACCINATION; RESPONSES; REVEALS; DEEP
Popis High-throughput sequencing analysis of hypermutating immunoglobulin (IG) repertoires remains a challenging task. Here we present a robust protocol for the full-length profiling of human and mouse IG repertoires. This protocol uses unique molecular identifiers (UMIs) introduced in the course of cDNA synthesis to control bottlenecks and to eliminate PCR and sequencing errors. Using asymmetric 400+100-nt paired-end Illumina sequencing and UMI-based assembly with the new version of the MIGEC software, the protocol allows up to 750-nt lengths to be sequenced in an almost error-free manner. This sequencing approach should also be applicable to various tasks beyond immune repertoire studies. In IG profiling, the achieved length of high-quality sequence covers the variable region of even the longest chains, along with the fragment of a constant region carrying information on the antibody isotype. The whole protocol, including preparation of cells and libraries, sequencing and data analysis, takes 5 to 6 d.
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