High-quality full-length immunoglobulin profiling with unique molecular barcoding
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14740%2F16%3A00088911" target="_blank" >RIV/00216224:14740/16:00088911 - isvavai.cz</a>
Result on the web
<a href="http://www.nature.com/nprot/journal/v11/n9/full/nprot.2016.093.html#access" target="_blank" >http://www.nature.com/nprot/journal/v11/n9/full/nprot.2016.093.html#access</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1038/nprot.2016.093" target="_blank" >10.1038/nprot.2016.093</a>
Alternative languages
Result language
angličtina
Original language name
High-quality full-length immunoglobulin profiling with unique molecular barcoding
Original language description
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.
Czech name
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Czech description
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Classification
Type
J<sub>x</sub> - Unclassified - Peer-reviewed scientific article (Jimp, Jsc and Jost)
CEP classification
CE - Biochemistry
OECD FORD branch
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Result continuities
Project
Result was created during the realization of more than one project. More information in the Projects tab.
Continuities
P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)
Others
Publication year
2016
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
NATURE PROTOCOLS
ISSN
1754-2189
e-ISSN
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Volume of the periodical
11
Issue of the periodical within the volume
9
Country of publishing house
GB - UNITED KINGDOM
Number of pages
18
Pages from-to
1599-1616
UT code for WoS article
000382164800004
EID of the result in the Scopus database
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