An Optimised Step-by-Step Protocol for Measuring Relative Telomere Length.

Autor:	Joglekar MV; Diabetes and Islet biology, NHMRC Clinical Trials Centre, Faculty of Medicine and Health, University of Sydney, Camperdown, NSW 2150, Australia., Satoor SN; Diabetes and Islet biology, NHMRC Clinical Trials Centre, Faculty of Medicine and Health, University of Sydney, Camperdown, NSW 2150, Australia.; DNA Sequencing Laboratory, National Centre for Cell Science, NCMR Campus, Sai Trinity Complex, Pashan, Pune 411 021, India., Wong WKM; Diabetes and Islet biology, NHMRC Clinical Trials Centre, Faculty of Medicine and Health, University of Sydney, Camperdown, NSW 2150, Australia., Cheng F; Department of Medicine & Therapeutics and Li Ka Shing Institute of Health Sciences, Chinese University of Hong Kong, Prince of Wales Hospital, Hong Kong, China., Ma RCW; Department of Medicine & Therapeutics and Li Ka Shing Institute of Health Sciences, Chinese University of Hong Kong, Prince of Wales Hospital, Hong Kong, China., Hardikar AA; Diabetes and Islet biology, NHMRC Clinical Trials Centre, Faculty of Medicine and Health, University of Sydney, Camperdown, NSW 2150, Australia.
Jazyk:	angličtina
Zdroj:	Methods and protocols [Methods Protoc] 2020 Apr 03; Vol. 3 (2). Date of Electronic Publication: 2020 Apr 03.
DOI:	10.3390/mps3020027
Abstrakt:	Telomeres represent the nucleotide repeat sequences at the ends of chromosomes and are essential for chromosome stability. They can shorten at each round of DNA replication mainly because of incomplete DNA synthesis of the lagging strand. Reduced relative telomere length is associated with aging and a range of disease states. Different methods such as terminal restriction fragment analysis, real-time quantitative PCR (qPCR) and fluorescence in situ hybridization are available to measure telomere length; however, the qPCR-based method is commonly used for large population-based studies. There are multiple variations across qPCR-based methods, including the choice of the single-copy gene, primer sequences, reagents, and data analysis methods in the different reported studies so far. Here, we provide a detailed step-by-step protocol that we have optimized and successfully tested in the hands of other users. This protocol will help researchers interested in measuring relative telomere lengths in cells or across larger clinical cohort/study samples to determine associations of telomere length with health and disease. Competing Interests: The authors declare no conflict of interest
Databáze:	MEDLINE
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