Preparing unbiased T cell receptor and antibody cDNA libraries for the deep next generation sequencing profiling

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14740%2F13%3A00081957" target="_blank" >RIV/00216224:14740/13:00081957 - isvavai.cz</a>

Výsledek na webu

<a href="http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3870325/" target="_blank" >http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3870325/</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.3389/fimmu.2013.00456" target="_blank" >10.3389/fimmu.2013.00456</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Preparing unbiased T cell receptor and antibody cDNA libraries for the deep next generation sequencing profiling

Popis výsledku v původním jazyce

High throughput sequencing has the power to reveal the nature of adaptive immunity as represented by the full complexity of T cell receptor (TCR) and antibody (IG) repertoires, but is at present severely compromised by the quantitative bias, bottlenecks,and accumulated errors that inevitably occur in the course of library preparation and sequencing. Here we report an optimized protocol for the unbiased preparation of TCR and IG cDNA libraries for high throughput sequencing, starting from thousands or millions of live cells in an investigated sample. Critical points to control are revealed, along with tips that allow researchers to minimize quantitative bias, accumulated errors, and cross sample contamination at each stage, and to enhance the subsequent bioinformatic analysis. The protocol is simple, reliable, and can be performed in 1 to 2 days.

Název v anglickém jazyce

Preparing unbiased T cell receptor and antibody cDNA libraries for the deep next generation sequencing profiling

Popis výsledku anglicky

High throughput sequencing has the power to reveal the nature of adaptive immunity as represented by the full complexity of T cell receptor (TCR) and antibody (IG) repertoires, but is at present severely compromised by the quantitative bias, bottlenecks,and accumulated errors that inevitably occur in the course of library preparation and sequencing. Here we report an optimized protocol for the unbiased preparation of TCR and IG cDNA libraries for high throughput sequencing, starting from thousands or millions of live cells in an investigated sample. Critical points to control are revealed, along with tips that allow researchers to minimize quantitative bias, accumulated errors, and cross sample contamination at each stage, and to enhance the subsequent bioinformatic analysis. The protocol is simple, reliable, and can be performed in 1 to 2 days.

Druh

J_x - Nezařazeno - Článek v odborném periodiku (Jimp, Jsc a Jost)

CEP obor

EB - Genetika a molekulární biologie

OECD FORD obor

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Projekt

<a href="/cs/project/ED1.1.00%2F02.0068" target="_blank" >ED1.1.00/02.0068: CEITEC - central european institute of technology</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2013

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Frontiers in Immunology

ISSN

1664-3224

e-ISSN

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Svazek periodika

4

Číslo periodika v rámci svazku

Dec

Stát vydavatele periodika

GB - Spojené království Velké Británie a Severního Irska

Počet stran výsledku

10

Strana od-do

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Kód UT WoS článku

000209374100444

EID výsledku v databázi Scopus

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