Validation of housekeeping genes as internal controls for studying biomarkers of endocrine-disrupting chemicals in disk abalone by real-time PCR

2011 ◽  
Vol 153 (3) ◽  
pp. 259-268 ◽  
Author(s):  
Qiang Wan ◽  
Ilson Whang ◽  
Cheol Young Choi ◽  
Jae-Seong Lee ◽  
Jehee Lee
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Endocrine Disrupting Chemicals ◽  
Housekeeping Genes ◽  
Internal Controls ◽  
Endocrine Disrupting

scholarly journals Beta-2 microglobulin a robust reference housekeeping genes for RNA expression normalization in real time PCR on human leukocytes

2019 ◽  
Author(s):  
Clara MILHEM ◽  
Céline INGELAERE ◽  
Olivier MORALES ◽  
Nadira DELHEM
Keyword(s):  
Gene Expression ◽  
Real Time ◽  
Real Time Pcr ◽  
Housekeeping Genes ◽  
Internal Controls ◽  
Rna Expression ◽  
Rt Pcr ◽  
Genomic Study ◽  
Beta 2 ◽  
Beta 2 Microglobulin

Abstract Background: Changes in gene expression are increasingly used to evaluate the effects of exposure to environmental agents. Housekeeping genes (Hk) are essential in these analyzes as internal controls to normalize expression levels assessed in real-time PCR (RT-PCR). Ideal Hk genes (i) are constitutively expressed; (ii) do not respond to external stimuli and (iii) show small or no variation between samples or from one assay to another. Previous studies indicate that some commonly used Hk genes, including glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and beta-actin, have differential expression in various cell lines. The objective of this study is to identify and validate the most appropriate housekeeping genes for RNA expression analysis of human primary peripheral blood mononuclear cells (PBMCs) in response to antigen like stimulation. Methods: The expression of the five most commonly used genes for a genomic study of lymphocytes was determined [HPRT, 18S, GAPDH, Ubiquitin C (UBC), beta-2 microglobulin (B2M)] on PBMCs, isolated from fresh blood, following activation with anti-CD3 and anti-CD28. This leads to the activation and proliferation of T lymphocytes in order to modulate the expression of specific genes. Results : Using reverse transcriptase RT-PCR protocol, we show that following activation only B2M remain unchanged in PBMCs. This result has been confirmes In contrast, 18S, HPRT and GAPDH show significant variation in PBMC gene expression. Conclusion: Although our results suggest that the relevance of Hk genes should be determined for each experimental condition, B2M appear to be excellent candidate as internal controls. Keywords : Housekeeping genes, RT-PCR, Human primary PBMC.

Validation of Housekeeping Genes as Internal Controls for the Study of the Effects of Microcystin-LR in Zebrafish by Real-Time PCR

Zebrafish ◽  
2018 ◽  
Vol 15 (5) ◽  
pp. 454-459 ◽  
Author(s):  
Mauricio da Silva Sopezki ◽  
José Maria Monserrat ◽  
João Sarkis Yunes ◽  
Juliano Zanette
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Housekeeping Genes ◽  
Internal Controls

scholarly journals Transcription of nanos-1 in Zebrafish Embryos is not Affected by Bisphenol A: Evaluated Using Quantitative Real-Time PCR

2019 ◽  
Vol 16 (1) ◽  
pp. 15-21
Author(s):  
Bridget Babich ◽  
George Roba ◽  
Siti Sarah Safura ◽  
Kevin Callahan ◽  
Edward Freeman
Keyword(s):  
Bisphenol A ◽  
Real Time ◽  
Germ Cells ◽  
Real Time Pcr ◽  
Target Gene ◽  
Primordial Germ Cells ◽  
Endocrine Disrupting Chemicals ◽  
Zebrafish Embryos ◽  
Quantitative Real Time Pcr ◽  
Endocrine Disrupting

The presence of primordial germ cells (PGCs) is crucial for proper gonad formation in zebrafish (Danio rerio). The many aspects of PGC migration that allow these cells to reach the proper location at the gonadal ridge include receptors, ligands, germ plasm components, and internal maintenance of PGCs. Any one of these factors could be affected by endocrine-disrupting chemicals (EDCs), which have been shown to alter the directed migration of these cells during early embryonic development. Based on recent research wherein the EDC bisphenol A (BPA) inhibited normal PGC migration, we have used the same dose of BPA to determine the impact of BPA on a gene central to proper germ cell migration. Zebrafish embryos were exposed to BPA, and the levels of the target gene nanos-1 were analyzed using quantitative real-time PCR (q-PCR). The target gene nanos-1 is a critically important germplasm component that allows for survival and proper migration of PGCs. The q-PCR results showed that BPA did not affect the transcription level of nanos-1 in zebrafish embryos. KEYWORDS: Zebrafish; Zebrafish Embryos; nanos-1; Primordial Germ Cells; PGC Migration; Gonad Development; Endocrine-Disrupting Chemicals; Bisphenol A; Sex Determination

scholarly journals Identification and Validation of Suitable Housekeeping Genes for Normalizing Quantitative Real-Time PCR Assays in Injured Peripheral Nerves

PLoS ONE ◽  
2014 ◽  
Vol 9 (8) ◽  
pp. e105601 ◽  
Author(s):  
Giovanna Gambarotta ◽  
Giulia Ronchi ◽  
Olivier Friard ◽  
Pantaleo Galletta ◽  
Isabelle Perroteau ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Peripheral Nerves ◽  
Housekeeping Genes ◽  
Quantitative Real Time Pcr ◽  
Pcr Assays

Free circulating nucleic acids in plasma and serum as a novel approach to the use of internal controls in real time PCR based detection

2014 ◽  
Vol 207 ◽  
pp. 133-137 ◽  
Author(s):  
Ersin Karataylı ◽  
Yasemin Çelik Altunoğlu ◽  
Senem Ceren Karataylı ◽  
Cihan Yurdaydın ◽  
A. Mithat Bozdayı
Keyword(s):  
Nucleic Acids ◽  
Real Time ◽  
Real Time Pcr ◽  
Internal Controls ◽  
Circulating Nucleic Acids ◽  
Novel Approach

scholarly journals Diurnal variation in opsin expression and common housekeeping genes necessitates comprehensive normalization methods for quantitative real‐time PCR analyses

2019 ◽  
Vol 19 (6) ◽  
pp. 1447-1460 ◽  
Author(s):  
Miranda R. Yourick ◽  
Benjamin A. Sandkam ◽  
William J. Gammerdinger ◽  
Daniel Escobar‐Camacho ◽  
Sri Pratima Nandamuri ◽  
...  
Keyword(s):  
Diurnal Variation ◽  
Real Time ◽  
Real Time Pcr ◽  
Housekeeping Genes ◽  
Quantitative Real Time Pcr ◽  
Opsin Expression ◽  
Normalization Methods
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