Multiplex preamplification of specific cDNA targets prior to gene expression analysis by TaqMan Arrays.

BMC Research Notes
Lourdes MengualAntonio Alcaraz

Abstract

An accurate gene expression quantification using TaqMan Arrays (TA) could be limited by the low RNA quantity obtained from some clinical samples. The novel cDNA preamplification system, the TaqMan PreAmp Master Mix kit (TPAMMK), enables a multiplex preamplification of cDNA targets and therefore, could provide a sufficient amount of specific amplicons for their posterior analysis on TA. A multiplex preamplification of 47 genes was performed in 22 samples prior to their analysis by TA, and relative gene expression levels of non-preamplified (NPA) and preamplified (PA) samples were compared. Overall, the mean cycle threshold (CT) decrement in the PA genes was 3.85 (ranging from 2.07 to 5.01). A high correlation (r) between the gene expression measurements of NPA and PA samples was found (mean r = 0.970, ranging from 0.937 to 0.994; p < 0.001 in all selected cases). High correlation coefficients between NPA and PA samples were also obtained in the analysis of genes from degraded RNA samples and/or low abundance expressed genes. We demonstrate that cDNA preamplification using the TPAMMK before TA analysis is a reliable approach to simultaneously measure gene expression of multiple targets in a single sample. Moreover, this procedure...Continue Reading

References

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Citations

Oct 2, 2012·BMC Genomics·Man RaoAlain Vignal
Jan 14, 2016·European Journal of Cancer : Official Journal for European Organization for Research and Treatment of Cancer (EORTC) [and] European Association for Cancer Research (EACR)·Maria J RibalAntonio Alcaraz
Feb 11, 2015·Schizophrenia Research·Greg C BristowMonsheel S Sodhi
Nov 8, 2011·Urology·Mercedes Marín-AguileraAntonio Alcaraz
May 21, 2013·International Journal of Cancer. Journal International Du Cancer·Lourdes MengualAntonio Alcaraz
Apr 19, 2015·BMC Molecular Biology·Vlasta KorenkováRobert Sjöback
Oct 6, 2018·PloS One·Annelore SacreasMinnie M Sarwal
Apr 22, 2020·The Plant Journal : for Cell and Molecular Biology·Youshi TazoeTsuyoshi Endo
Feb 18, 2017·Oncotarget·Mercedes Ingelmo-TorresLourdes Mengual
Apr 22, 2010·Clinical Cancer Research : an Official Journal of the American Association for Cancer Research·Lourdes MengualAntonio Alcaraz

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Methods Mentioned

BETA
PCR

Software Mentioned

PCR
SDS
DART

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