Dijital PZR ve kullanım alanları

Year 2016, Volume: 73 Issue: 2, 183 - 198, 01.06.2016

Ahmet Çarhan Elif Ercan Tuğba Yalçınkaya

Abstract

Polimeraz zincir reaksiyonu PZR , DNA kompleks havuzundan spesifik bir DNA parçasını çoğaltmaya olanak sağlayan basit, etkin ve özellikle moleküler biyoloji alanında yaygın olarak kullanılan enzimatik bir tekniktir. PZR tekniğinin keşfinden günümüze kadar gelişen teknolojiyle birlikte pek çok PZR tekniği ortaya çıkmıştır. Bu tekniklerden bazıları Gerçek Zamanlı PZR, Kantitatif PZR, Ters Transkriptaz PZR, Nested PZR ve Multipleks PZR dir. Günümüze kadar geliştirilmiş PZR tekniklerinin çeşitli zorlukları yüzünden PZR tekniklerinin kullanım alanını genişletmek ve daha ileri seviye PZR teknikleri bulmak araştırmacıların birinci önceliği haline gelmiştir. Nadir mutasyonların, kopya sayısı varyasyonlarındaki ufak değişikliklerin, gen ifadesi değişiklikleri arasındaki farklılıkların veya metilasyon durumunun değerlendirilmesine izin veren yeni bir yöntem olarak dijital PZR dPZR teknolojisi geliştirilmiştir. Dijital PZR, DNA kopya sayısının hassas ölçümü için PZR bazlı yeni bir tekniktir ve örnekleri az sayıda seyrelterek çok sayıda PZR gerçekleştirmeye olanak sağlar ve her birinde ayrı ayrı PZR gerçekleşen çok sayıda küçük bölümlere sahiptir. Aynı zamanda dPZR; şu an çok az miktardaki genetik materyalin miktarını dakikalar içinde tespit etme performansıyla birçok nicel yöntemleri geride bırakmıştır. Tek molekülü sayma stratejisi sayesinde bu yöntem yüksek hassasiyet gösterebilmektedir. Güvenilirlik ve tekrarlanabilirlik düzeyi oldukça yüksek bir yöntem olmasıyla da dikkat çekmektedir. Bu derleme, digital PZR yönteminin günümüze kadar olan süreçteki gelişimine ve ilerleyen günlerde çözüm bulunması gereken eksik yönlerine dikkat çekmeyi amaçlamaktadır

Keywords

Dijital PZR, Nadir Mutasyon, dMIQE Kuralları

Digital PCR and applications

Abstract

Polymerase chain reaction PCR , is a simple, effective and widely used enzymatic technic especially in the field of molecular biology which provides the opportunity to amplify a specific DNA fragment from DNA complex pool. Several PCR techniques such as realtime PCR, quantitative and qualitative PCR, Reverse Transcriptase PCR, Nested PCR and multiplex PCR have been developed since the first invention of PCR. Due to the various difficulties of the PCR techniques developed so far, widening the application fields and finding out more advanced level of PCR techniques have become the first priority of the researchers. Digital PCR dPCR technology has been developed as a new method to permit the evaluation of the small changes in the copy number variations of the rare mutations, differences between the gene expression changes or state of methylation. Digital PCR is a PCR based new technique for the sensitive measurement of number of DNA copies, it provides opportunity to do large number of PCR with a few number of sample dilution and it has so many small compartments where seperate PCRs are executed in each. At the same time, dPCR leaves some of the quantitative methods behind with the performance of determining the quantity of small amount of genetic material within seconds. This method shows high

Keywords

Digital PCR, Rare Mutation, dMIQE rules

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