oru.sePublikasjoner
Endre søk
RefereraExporteraLink to record
Permanent link

Direct link
Referera
Referensformat
  • apa
  • harvard1
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Annet format
Fler format
Språk
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Annet språk
Fler språk
Utmatningsformat
  • html
  • text
  • asciidoc
  • rtf
Volume-related inhibitors standardization for reverse transcription quantitative polymerase chain reaction experiments
University of Kentucky, Lexington KY, United States .ORCID-id: 0000-0002-5322-4150
Vise andre og tillknytning
2012 (engelsk)Konferansepaper, Poster (with or without abstract) (Fagfellevurdert)
Abstract [en]

A large part of the reliability of reverse transcription quantitative polymerase chain reaction (RT-qPCR) data depends on technical variations. Such variations are mainly attributable to the reverse transcription step. Standardization is a key factor in decreasing the intersample variability. However, an ideal standardization is not always possible, and compromises must be found. Due to technical requirements, the current consensus is that a constant amount of total RNA should be used for the RT step (CA-RT). Because RNA isolation yields are variable, such a practice requires the use of variable volumes of nucleic acid extracts in RT reaction. We demonstrate that some RNA extracts contain both exogenous and endogenous inhibitors. These inhibitors induce a decreased RT efficiency that significantly impairs the reliability of RT-qPCR data. Conversely, these inhibitors have slight effects on the qPCR step. To overcome such drawbacks, we proposed to carry out the RT reaction with a constant volume of RNA extract by preserving a constant RNA amount through the supplementation of yeast transfer RNA (CV-RT). We show that CV-RT, compared with the usual CA-RT, allows us to decrease the RT-qPCR variability induced by intersample differences. Such a decrease is a prerequisite for the reliability of messenger RNA quantification.

sted, utgiver, år, opplag, sider
2012.
HSV kategori
Identifikatorer
URN: urn:nbn:se:oru:diva-66331OAI: oai:DiVA.org:oru-66331DiVA, id: diva2:1194902
Konferanse
Congress of Genomics Research, Boston, USA, April 19-20, 2012
Tilgjengelig fra: 2018-04-04 Laget: 2018-04-04 Sist oppdatert: 2018-05-15bibliografisk kontrollert

Open Access i DiVA

Fulltekst mangler i DiVA

Personposter BETA

Chaillou, Thomas

Søk i DiVA

Av forfatter/redaktør
Chaillou, Thomas

Søk utenfor DiVA

GoogleGoogle Scholar

urn-nbn

Altmetric

urn-nbn
Totalt: 108 treff
RefereraExporteraLink to record
Permanent link

Direct link
Referera
Referensformat
  • apa
  • harvard1
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Annet format
Fler format
Språk
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Annet språk
Fler språk
Utmatningsformat
  • html
  • text
  • asciidoc
  • rtf