Identification and validation of reference genes for qPCR in the terrestrial gastropod Cepaea nemoralis

PloS One
Susanne AffenzellerDaniel John Jackson

Abstract

Identifying and understanding mechanisms that generate phenotypic diversity is a fundamental goal of evolutionary biology. With a diversity of pigmented shell morphotypes governed by Mendelian patterns of inheritance, the common grove snail Cepaea nemoralis (Linnaeus, 1758) has been a model for evolutionary biologists and population geneticists for decades. However, the genetic mechanisms by which C. nemoralis generates this pigmented shell diversity remain unknown. An important first step in investigating this pigmentation pattern is to establish a set of validated reference genes for differential gene expression assays. Here we have evaluated eleven candidate genes for reverse transcription quantitative polymerase chain reaction (qPCR) in C. nemoralis. Five of these were housekeeping genes traditionally employed as qPCR reference genes in other species, while six alternative genes were selected de novo from C. nemoralis transcriptome data based on the stability of their expression levels. We tested all eleven candidates for expression stability in four sub-adult tissues of C. nemoralis: pigmented mantle, unpigmented mantle, head and foot. We find that two commonly employed housekeeping genes (alpha tubulin, glyceraldehyde 3-p...Continue Reading

References

Jan 1, 1959·Cold Spring Harbor Symposia on Quantitative Biology·M LAMOTTE
Jan 7, 2004·Biochemical and Biophysical Research Communications·Aleksandar RadonićAndreas Nitsche
Dec 1, 1950·Heredity·A J CAIN, P M SHEPPARD
Jul 31, 2004·BioTechniques·Keertan DhedaAlimuddin Zumla
Mar 24, 2007·Bioinformatics·Triinu Koressaar, Maido Remm
Mar 28, 2008·PloS One·Lieven ThorrezFrans Schuit
Feb 25, 2009·Nucleic Acids Research·J M RuijterA F M Moorman
Dec 21, 2010·Comparative Biochemistry and Physiology. Toxicology & Pharmacology : CBP·Qiang WanJehee Lee
Feb 2, 2012·Environmental Science and Pollution Research International·Elena Cubero-LeonJeanette M Rotchell
Jun 26, 2012·Nucleic Acids Research·Andreas UntergasserSteven G Rozen
Apr 4, 2014·Bioinformatics·Anthony M BolgerBjoern Usadel
Apr 5, 2014·Current Biology : CB·Tanja SchwanderLaurent Keller
Apr 15, 2016·Biomolecular Detection and Quantification·Jan M RuijterMaurice J B van den Hoff
Aug 5, 2017·Science·Innes C CuthillTim Caro
Sep 28, 2017·PeerJ·Jesse KerkvlietKen Kraaijeveld

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Citations

Feb 14, 2020·Scientific Reports·Susanne AffenzellerDaniel J Jackson

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Datasets Mentioned

BETA
MH035488
MH035489
MH035491
MH035493
MH035498
MH035490
MH035492
MH035494
MH035495
MH035496

Methods Mentioned

BETA
electrophoresis
PCR

Software Mentioned

CLC
LinRegPCR
Primer3
NormFinder
Gene Q
tBLASTx
tran
geNorm
Gene
Trinity

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