Technical aspects and recommendations for single-cell qPCR

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F86652036%3A_____%2F18%3A00503182" target="_blank" >RIV/86652036:_____/18:00503182 - isvavai.cz</a>

Výsledek na webu

<a href="http://dx.doi.org/10.1016/j.mam.2017.07.004" target="_blank" >http://dx.doi.org/10.1016/j.mam.2017.07.004</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1016/j.mam.2017.07.004" target="_blank" >10.1016/j.mam.2017.07.004</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Technical aspects and recommendations for single-cell qPCR

Popis výsledku v původním jazyce

Single cells are basic physiological and biological units that can function individually as well as in groups in tissues and organs. It is central to identify, characterize and profile single cells at molecular level to be able to distinguish different kinds, to understand their functions and determine how they interact with each other. During the last decade several technologies for single-cell profiling have been developed and used in various applications, revealing many novel findings. Quantitative PCR (qPCR) is one of the most developed methods for single-cell profiling that can be used to interrogate several analytes, including DNA, RNA and protein. Single-cell qPCR has the potential to become routine methodology but the technique is still challenging, as it involves several experimental steps and few molecules are handled. Here, we discuss technical aspects and provide recommendation for single-cell qPCR analysis. The workflow includes experimental design, sample preparation, single-cell collection, direct lysis, reverse transcription, preamplification, qPCR and data analysis. Detailed reporting and sharing of experimental details and data will promote further development and make validation studies possible. Efforts aiming to standardize single-cell qPCR open up means to move single-cell analysis from specialized research settings to standard research laboratories. (C) 2017 Elsevier Ltd. All rights reserved.

Název v anglickém jazyce

Technical aspects and recommendations for single-cell qPCR

Popis výsledku anglicky

Single cells are basic physiological and biological units that can function individually as well as in groups in tissues and organs. It is central to identify, characterize and profile single cells at molecular level to be able to distinguish different kinds, to understand their functions and determine how they interact with each other. During the last decade several technologies for single-cell profiling have been developed and used in various applications, revealing many novel findings. Quantitative PCR (qPCR) is one of the most developed methods for single-cell profiling that can be used to interrogate several analytes, including DNA, RNA and protein. Single-cell qPCR has the potential to become routine methodology but the technique is still challenging, as it involves several experimental steps and few molecules are handled. Here, we discuss technical aspects and provide recommendation for single-cell qPCR analysis. The workflow includes experimental design, sample preparation, single-cell collection, direct lysis, reverse transcription, preamplification, qPCR and data analysis. Detailed reporting and sharing of experimental details and data will promote further development and make validation studies possible. Efforts aiming to standardize single-cell qPCR open up means to move single-cell analysis from specialized research settings to standard research laboratories. (C) 2017 Elsevier Ltd. All rights reserved.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10608 - Biochemistry and molecular biology

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

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

Rok uplatnění

2018

Kód důvěrnosti údajů

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

Název periodika

Molecular Aspects of Medicine

ISSN

0098-2997

e-ISSN

—

Svazek periodika

59

Číslo periodika v rámci svazku

FEB 2018

Stát vydavatele periodika

NL - Nizozemsko

Počet stran výsledku

8

Strana od-do

28-35

Kód UT WoS článku

000423056300003

EID výsledku v databázi Scopus

2-s2.0-85026450067