An Optimised Step-by-Step Protocol for Measuring Relative Telomere Length
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...
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doaj-9f1db1583dad4cb896464b58cef02c092020-11-25T02:27:11ZengMDPI AGMethods and Protocols2409-92792020-04-013272710.3390/mps3020027An Optimised Step-by-Step Protocol for Measuring Relative Telomere LengthMugdha V. Joglekar0Sarang N. Satoor1Wilson K.M. Wong2Feifei Cheng3Ronald C.W. Ma4Anandwardhan A. Hardikar5Diabetes and Islet biology, NHMRC Clinical Trials Centre, Faculty of Medicine and Health, University of Sydney, Camperdown, NSW 2150, AustraliaDiabetes and Islet biology, NHMRC Clinical Trials Centre, Faculty of Medicine and Health, University of Sydney, Camperdown, NSW 2150, AustraliaDiabetes and Islet biology, NHMRC Clinical Trials Centre, Faculty of Medicine and Health, University of Sydney, Camperdown, NSW 2150, AustraliaDepartment of Medicine & Therapeutics and Li Ka Shing Institute of Health Sciences, Chinese University of Hong Kong, Prince of Wales Hospital, Hong Kong, ChinaDepartment of Medicine & Therapeutics and Li Ka Shing Institute of Health Sciences, Chinese University of Hong Kong, Prince of Wales Hospital, Hong Kong, ChinaDiabetes and Islet biology, NHMRC Clinical Trials Centre, Faculty of Medicine and Health, University of Sydney, Camperdown, NSW 2150, AustraliaTelomeres 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.https://www.mdpi.com/2409-9279/3/2/27Telomeresreal-time PCRrelative telomere lengthcellsDNA |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Mugdha V. Joglekar Sarang N. Satoor Wilson K.M. Wong Feifei Cheng Ronald C.W. Ma Anandwardhan A. Hardikar |
spellingShingle |
Mugdha V. Joglekar Sarang N. Satoor Wilson K.M. Wong Feifei Cheng Ronald C.W. Ma Anandwardhan A. Hardikar An Optimised Step-by-Step Protocol for Measuring Relative Telomere Length Methods and Protocols Telomeres real-time PCR relative telomere length cells DNA |
author_facet |
Mugdha V. Joglekar Sarang N. Satoor Wilson K.M. Wong Feifei Cheng Ronald C.W. Ma Anandwardhan A. Hardikar |
author_sort |
Mugdha V. Joglekar |
title |
An Optimised Step-by-Step Protocol for Measuring Relative Telomere Length |
title_short |
An Optimised Step-by-Step Protocol for Measuring Relative Telomere Length |
title_full |
An Optimised Step-by-Step Protocol for Measuring Relative Telomere Length |
title_fullStr |
An Optimised Step-by-Step Protocol for Measuring Relative Telomere Length |
title_full_unstemmed |
An Optimised Step-by-Step Protocol for Measuring Relative Telomere Length |
title_sort |
optimised step-by-step protocol for measuring relative telomere length |
publisher |
MDPI AG |
series |
Methods and Protocols |
issn |
2409-9279 |
publishDate |
2020-04-01 |
description |
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. |
topic |
Telomeres real-time PCR relative telomere length cells DNA |
url |
https://www.mdpi.com/2409-9279/3/2/27 |
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