Sethi Itty, Bhat Gh Rasool, Kumar Rakesh, Rai Ekta, Sharma Swarkar
Human Genetics Research Group, School of Biotechnology, Shri Mata Vaishno Devi University, Katra 182320, India.
Human Genetics Research Group, School of Biotechnology, Shri Mata Vaishno Devi University, Katra 182320, India.
Gene. 2021 Jan 30;767:145178. doi: 10.1016/j.gene.2020.145178. Epub 2020 Sep 30.
Telomeres are highly repetitive regions capping the chromosomes and composed of multiple units of hexa-nucleotides, TTAGGG, making their quantification difficult. Most of the methods developed to estimate telomeres are extensively cumbersome or expensive. The quantitative polymerase chain reaction (qPCR) based assay is relatively easy and cheaper method that applies SyBr Green dye chemistry to measure telomere length. SyBr Green dye fluoresces after intercalation into the double stranded DNA (dsDNA), thus detection of unspecific products has been a limitation as it may affect quantitation of telomeres. To overcome this limitation of SyBr Green dye, we developed a dual labeled fluorescence probe based quantitative polymerase chain reaction (qPCR) to measure the telomere length. This highly efficient, yet cost effective and easy method, utilizes a probe that targets primarily the telomeric DNA and this increases accuracy of an existing qPCR method.
端粒是位于染色体末端的高度重复区域，由多个六核苷酸单元TTAGGG组成，这使得对其进行定量分析具有一定难度。大多数用于估计端粒的方法都非常繁琐或昂贵。基于定量聚合酶链反应（qPCR）的检测方法相对简单且成本较低，该方法应用SYBR Green染料化学来测量端粒长度。SYBR Green染料在嵌入双链DNA（dsDNA）后会发出荧光，因此非特异性产物的检测一直是一个限制因素，因为它可能会影响端粒的定量分析。为了克服SYBR Green染料的这一局限性，我们开发了一种基于双标记荧光探针的定量聚合酶链反应（qPCR）来测量端粒长度。这种高效、经济且简便的方法利用了一种主要靶向端粒DNA的探针，从而提高了现有qPCR方法的准确性。
