Identification and Validation of Reference Genes for RT-qPCR Normalization in Nauphoeta cinerea (Olivier, 1789) (Blattodea, Blaberidae)

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dc.contributor.authorÖzcan, Kübra
dc.contributor.authorPektaş, Ayşe Nur
dc.contributor.authorBerk, Şeyda
dc.date.accessioned2025-05-04T16:21:08Z
dc.date.available2025-05-04T16:21:08Z
dc.date.issued2022
dc.departmentSivas Cumhuriyet Üniversitesi
dc.description.abstractQuantitative RT-PCR (q-RT-PCR) is a powerful tool that allows large-scale analysis of very small changes in gene expression. For the calculation of gene expression, such as the delta-delta Ct method, different PCR primer efficiencies (E) may affect the result, as PCR primer yields are assumed to be comparable for the gene of interest and housekeeping gene. Therefore, identification of a suitable reference gene for data normalization is an important step in the development of qPCR assays. Furthermore, accurate and reliable results depend on the use of stable reference genes for normalization. The aim of the current study is the identification and validation of a set of six housekeeping genes (GADPH, RPS18, α-TUB, EF1α, ArgK, and ACTB) in cockroach species Nauphoeta cinerea adults using five different algorithms (ΔCt method, Bestkeeper, geNorm, Normfinder and RefFinder) to evaluate the stability of selected reference genes expression. Our results show that α-Tub use provides accurate normalization of gene expression levels in N. cinerea adults. In addition, since the GADPH is selected as the second most stable reference gene, GADPH can be also used for transcript analysis N. cinerea adults. Our study also showed that ACTB (β-actin) should not be used for normalizing transcript levels when examining N. cinerea adults. Additionally, validation studies for reference genes in cockroaches are very few (only one) in the literature. Therefore, the results highlight the need for validation of reference genes under biotic and abiotic conditions in q-RT-PCR studies in cockroaches.
dc.description.abstract[No abstract available]
dc.identifier.doi10.30516/bilgesci.1067570
dc.identifier.doihttps://doi.org/10.30516/bilgesci.1067570
dc.identifier.endpage72
dc.identifier.issn2651-401X
dc.identifier.issn2651-4028
dc.identifier.issue1
dc.identifier.startpage62
dc.identifier.urihttps://hdl.handle.net/20.500.12418/31605
dc.identifier.volume6
dc.language.isoen
dc.publisherKutbilge Akademisyenler Derneği
dc.relation.ispartofBilge International Journal of Science and Technology Research
dc.relation.publicationcategoryMakale - Ulusal Hakemli Dergi - Kurum Öğretim Elemanı
dc.rightsinfo:eu-repo/semantics/openAccess
dc.snmzKA_DergiPark_20250504
dc.subjectRT-qPCR
dc.subjectreference genes
dc.subjectvalidation
dc.subjectnormalization
dc.subjectNauphoeta cinerea
dc.titleIdentification and Validation of Reference Genes for RT-qPCR Normalization in Nauphoeta cinerea (Olivier, 1789) (Blattodea, Blaberidae)
dc.typeResearch Article

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