SavedAdd your journal

Journals / Cumhuriyet Science Journal / 2022 / Cilt: 43 - Sayı: 2

Selection and Validation of Potential Reference Genes for Quantitative Real-Time PCR Analysis in Blaptica Dubia (Serville, 1838) (Blattidae, Blaberidae)

Emin Ufuk KARAKAŞ
Ayşe Nur PEKTAŞ
Şeyda BERK
Pages
176–182
Publication Date
2022-10-12
DOI
—
🏛️Bibliographic metadata and full text access for this article are provided via DergiPark.
DergiPark Record ↗
✨

AI Summary & Key Takeaways

AI Insight
⚡ TL;DR(Too Long; Didn't Read)

In addition, since there are few validation studies for reference genes in cockroaches in the literature, it is considered that it would be beneficial to increase the number of studies related with RT-qPCR on the reference genes validation under biotic and abiotic conditions in cockroaches.

📌 3 Key Takeaways

🎯
Objective & Hypothesis:Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) is an effective, reproducible, and dependable method for evaluating and targeting expression of genes.
📊
Key Findings & Data:The purpose of this study was to identify and validate six housekeeping genes (GADPH, RPS18, α-TUB, EF1α, ArgK and ACTB) in adults of cockroach species Blaptica dubia employing five different algorithms (geNorm, Bestkeeper, Normfinder, ΔCt method and RefFinder) to assess putative housekeeping gene expression stability.
💡
Conclusion & Impact:In addition, since there are few validation studies for reference genes in cockroaches in the literature, it is considered that it would be beneficial to increase the number of studies related with RT-qPCR on the reference genes validation under biotic and abiotic conditions in cockroaches.
🔬Design:Research Article
📍Scope:Comprehensive Sample
⚙️Variables:Domain Parameters

Abstract

Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) is an effective, reproducible, and dependable method for evaluating and targeting expression of genes. It is very important to normalize according to stably expressed housekeeping genes in order to facilitating gene expression studies and to acquire exact and meaningful results. The purpose of this study was to identify and validate six housekeeping genes (GADPH, RPS18, α-TUB, EF1α, ArgK and ACTB) in adults of cockroach species Blaptica dubia employing five different algorithms (geNorm, Bestkeeper, Normfinder, ΔCt method and RefFinder) to assess putative housekeeping gene expression stability. Our study also showed that the geNorm, Normfinder ΔCt method and RefFinder algorithms identified GADPH as the most stable housekeeping gene in B. dubia adults. Additioanlly, RPS18 was suggested as the most stable gene by GeNorm and BestKeeeper. ACTB has been shown to be by far the least stable of all algorithms. In addition, since there are few validation studies for reference genes in cockroaches in the literature, it is considered that it would be beneficial to increase the number of studies related with RT-qPCR on the reference genes validation under biotic and abiotic conditions in cockroaches.

🔖

Cite This Article

KARAKAŞ, E. U., PEKTAŞ, A. N., BERK, Ş. (2022). Selection and Validation of Potential Reference Genes for Quantitative Real-Time PCR Analysis in Blaptica Dubia (Serville, 1838) (Blattidae, Blaberidae). Cumhuriyet Science Journal, Cilt: 43 - Sayı: 2, 176–182. https://ajindex.com/articles/477505

📄 Full Text PDF Viewer

📄

PDF görüntüleyici hazırlanıyor...

References (12)

  1. Hudson M.E., Sequencing breakthroughs for genomic ecology and evolutionary biology, Mol. Ecol. Resour., 8 (2008) 3-17
  2. Li R., Xie W., Wang S., Wu Q., Yang N., Yang X., Pan H., Zhou X., Bai L., Xu B., Zhou X., Zhang Y., Reference gene selection for qRT-PCR analysis in the sweetpotato whitefly, Bemisia tabaci (Hemiptera: Aleyrodidae), PLoS One, 8 (2013) e53006
  3. Lourenço A.P., Mackert A., Cristino A.d.S., Simões Z.L.P., Validation of reference genes for gene expression studies in the honey bee, Apis mellifera, by quantitative real-time RT-PCR, Apidologie, 39 (2008) 372-385
  4. Vandesompele J., De Preter K., Pattyn F., Poppe B., Van Roy N., De Paepe A., Speleman F., Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes, Genome biology, 3 (2002) Research0034
  5. Strube C., Buschbaum S., Wolken S., Schnieder T., Evaluation of reference genes for quantitative real-time PCR to investigate protein disulfide isomerase transcription pattern in the bovine lungworm Dictyocaulus viviparus, Gene, 425 (2008) 36-43
  6. Guo J., Ling H., Wu Q., Xu L., Que Y., The choice of reference genes for assessing gene expression in sugarcane under salinity and drought stresses, Scientific Reports, 4 (2014) 7042
  7. Andersen C.L., Jensen J.L., Ørntoft T.F., Normalization of real-time quantitative reverse transcription-PCR data: a model-based variance estimation approach to identify genes suited for normalization, applied to bladder and colon cancer data sets, Cancer Research, 64 (2004) 5245-50
  8. Van Hiel M.B., Van Wielendaele P., Temmerman L., Van Soest S., Vuerinckx K., Huybrechts R., Broeck J.V., Simonet G., Identification and validation of housekeeping genes in brains of the desert locust Schistocerca gregaria under different developmental conditions, BMC Mol Biol., 10 (2009) 56
  9. Pitino M., Coleman A.D., Maffei M.E., Ridout C.J., Hogenhout S.A., Silencing of aphid genes by dsRNA feeding from plants, PLoS One, 6 (2011) e25709
  10. Fu W., Xie W., Zhang Z., Wang S., Wu Q., Liu Y., Zhou X., Zhou X., Zhang Y., Exploring valid reference genes for quantitative realtime PCR analysis in Plutella xylostella (Lepidoptera: Plutellidae), Int. J. Biol. Sci., 9 (2013) 792-802
  11. Berk S., and Pektas A., Selection and Validation of Reference Genes for Quantitative Real-time PCR in the Mealworm Beetle, Tenebrio molitorL. (Coleoptera: Tenebrionidae), IOSR-JAVS., 13 (2020) 44-50
  12. Ladror D.T., Frey B.L., Scal