Kantitatif RT-PCR (RT-qPCR) Uygulamalarında Kullanılan RNA’ların Kalite Kontrolleri ve Önemi

Year 2020, Volume: 17 Issue: 3, 334 - 341, 01.12.2020

Ali Osman Turgut Özgecan Korkmaz Ağaoğlu

https://doi.org/10.32707/ercivet.828871

Abstract

RNA kalitesi, RT-qPCR performansı ve güvenilirliği açısından dikkat edilmesi gereken önemli bir husustur. RT-qPCR çalışmalarında RNA kalite değerlendirilmesine dikkat edilmemesi, yapılan bilimsel çalışmaların güvenilirliği açı-sından önemli bir risk oluşturmaktadır. Bu noktada, RT-qPCR çalışmasının her aşamasında gerekli standartlara uyul-ması ve RNA kalite değerlendirmesinin mutlaka yapılması gerekmektedir. RNA kalitesinin çalışmanın her aşamasında yüksek tutulması, RT-qPCR verimliliğinin ve elde edilen sonuçların güvenilirliğinin artırılmasına olanak tanımaktadır. Yapılan çok sayıda çalışma RNA kalitesinin RT-qPCR sonuçları üzerine olan etkisini ve RNA kalite değerlendirmesinin gerekliliğini ortaya koymaktadır. Bu derlemede, RNA kalitesi, RNA kalite değerlendirmesine kullanılan yöntemler ve RNA kalitesinin RT-qPCR performansı üzerine etkilerinden bahsedilmiştir.

Keywords

Gen ekspresyonu, MIQE, RNA kalitesi, RT-qPCR

Quality Controls and Importance of RNAs Used in Quantitative RT-PCR (RT-qPCR) Applications

Abstract

RNA quality is an important consideration for RT-qPCR performance and reliability. The lack of attention to RNA quality assessment poses a significant risk for the reliability of scientific studies in RT-qPCR studies. At this point, the required standards must be followed, and RNA quality assessment must be performed at every stage of the RT-qPCR study. Maintaining high RNA quality at each stage of the study enables to increase efficiency of RT-qPCR and reliability of the results. Numerous studies have demonstrated the effect of RNA quality on RT-qPCR results and the need for RNA quality assessment. In this review; RNA quality, methods used for RNA quality assessment and the ef-fects of RNA quality on RT-qPCR performance are discussed.

Keywords

Gene expression, MIQE, RNA quality, RT-qPCR

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