Validation of quantitative polymerase chain reaction with Southern blot method for telomere length analysis

Tarik, Mohamad; Sachdev, Harshpal Singh; Tandon, Nikhil; Roy, Ambuj; Bhargava, Santosh K.; Pandey, Ravindra Mohan

doi:10.4155/fsoa-2017-0115

Cited by 16 publications

(15 citation statements)

References 32 publications

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“…The length of telomeres can be thought as the individual’s biological clock. There are different strategies for determining telomere length, including terminal restriction fragment (TRF), Southern blotting (SB) analysis, quantitative polymerase chain reaction (PCR), and flow-fluorescence in situ hybridization (FISH) (Tarik et al, 2018). TRF analysis is the oldest approach for determining telomere length.…”

Section: Telomeres Telomerase Activity and Telomere Lengthmentioning

confidence: 99%

The Alteration of Subtelomeric DNA Methylation in Aging-Related Diseases

Duan

2019

Front. Genet.

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The telomere is located at the end of the chromosome and consists of a non-coding, repetitive DNA sequence. As the cell divides, the length of telomere gradually decreases. A very short telomere can terminate mitosis, and thus telomere length becomes a hallmark of cellular aging. The 500 kb region of each autosomal arm terminal is the so-called subtelomeric region. Both telomere and subtelomere have high-density DNA repeats. Telomeres do not contain genes or CpG sequences, while subtelomeres contain small amounts of genes and high-density CpG sequences, and DNA methylation often occurs in subtelomeres. Previous studies have shown that aberrant methylation of subtelomeric DNA exists in many diseases, and it has a certain effect on the regulation of telomere length. In this review, we focus on the correlation between subtelomeric DNA methylation and aging-related diseases. We also summarize the relationship between subtelomeric methylation and telomere length in different diseases.

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Section: Telomeres Telomerase Activity and Telomere Lengthmentioning

confidence: 99%

The Alteration of Subtelomeric DNA Methylation in Aging-Related Diseases

Duan

2019

Front. Genet.

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“…One weakness of our study is that, we measured RTL by RT-PCR and not aTL by Southern blotting, which is the gold standard but difficult to perform in large number of samples. We did carry out a comparative analysis of RTL by RT-PCR and Southern blot in around 100 samples to validate our measurement and found a good agreement between the two methods (Tarik et al, 2018). Our study had several strengths.…”

Section: Conditional Bmi Gain and Telomere Lengthmentioning

confidence: 79%

“…RTL was determined by qPCR after extracting DNA from stored samples from Phase 6 and freshly collected samples from Phase 7. To validate the qPCR method, absolute telomere length (aTL) was measured by Southern blot assay in a subset of 94 samples as previously described (Tarik et al, ).…”

Section: Methodsmentioning

confidence: 99%

Association of birth outcomes and postnatal growth with adult leukocyte telomere length: Data from New Delhi Birth Cohort

Tarik

Sinha

Sachdev

et al. 2019

Maternal & Child Nutrition

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Born small for gestational age due to undernutrition in utero and subsequent catch‐up growth is associated with risk of developing chronic diseases in adulthood. Telomere length has been shown to be a predictor of these age‐related diseases and may be a link between birth size, a surrogate for foetal undernutrition, and adult chronic diseases. We assessed the relationship of leukocyte telomere length in adult life with birth outcomes and serial change in body mass index (BMI) from birth to adulthood. Leukocyte relative telomere length (RTL) was measured by MMqPCR in 1,309 subjects from New Delhi Birth Cohort who participated in two phases of the study between 2006–2009 (Phase 6) and 2012–2015 (Phase 7) at a mean age of 39.08 (±3.29), and its association with birth outcomes and conditional BMI gain at 2, 11, and 29 years was assessed in a mixed regression model. We did not find any significant association of RTL with body size at birth including birthweight, birth length, and birth BMI. Gestational age was positively associated with RTL (P = .017, multivariate model: P = .039). Conditional BMI gain at 2 and 11 years was not associated with RTL. BMI gain at 29 year was negatively associated with RTL in multivariate model (P = .015). Born small for gestational age was not associated with RTL in adulthood. Leukocyte telomere attrition was observed in those born before 37 weeks of gestational age as well as in those who gained weight as adults, which may predispose to chronic diseases.

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“…Pipetting errors can significantly affect the results and were partly overcome with an improved monochrome multiplex qPCR (mmqPCR) allowing the signals from the telomere amplicons and control amplicons to be collected simultaneously from the same well (Cawthon, 2009 ). Their main disadvantage is high intra- and inter-assay variability with reported coefficients of variation between 2 and 28% (Cawthon, 2002 , 2009 ; Martin-Ruiz et al, 2015 ; Tarik et al, 2018 ). Single Telomere Length Analysis (STELA) is a PCR based technique combined with Southern blot that measures TL on individual chromosomes (Baird et al, 2003 ).…”

Section: Telomere Biology In Human and Measurement Of Telomere Lengthmentioning

confidence: 99%

Telomere Attrition in Neurodegenerative Disorders

Levstek

Kozjek

Dolžan

et al. 2020

Front. Cell. Neurosci.

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Telomere attrition is increased in various disorders and is therefore a potential biomarker for diagnosis and/or prognosis of these disorders. The contribution of telomere attrition in the pathogenesis of neurodegenerative disorders is yet to be fully elucidated. We are reviewing the current knowledge regarding the telomere biology in two common neurodegenerative disorders, Alzheimer’s disease (AD), and Parkinson’s disease (PD). Furthermore, we are discussing future prospective of telomere research in these disorders. The majority of studies reported consistent evidence of the accelerated telomere attrition in AD patients, possibly in association with elevated oxidative stress levels. On the other hand in PD, various studies reported contradictory evidence regarding telomere attrition. Consequently, due to the low specificity and sensitivity, the clinical benefit of telomere length as a biomarker of neurodegenerative disease development and progression is not yet recognized. Nevertheless, longitudinal studies in large carefully selected cohorts might provide further elucidation of the complex involvement of the telomeres in the pathogenesis of neurodegenerative diseases. Telomere length maintenance is a complex process characterized by environmental, genetic, and epigenetic determinants. Thus, in addition to the selection of the study cohort, also the selection of analytical methods and types of biological samples for evaluation of the telomere attrition is of utmost importance.

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Validation of quantitative polymerase chain reaction with Southern blot method for telomere length analysis

Cited by 16 publications

References 32 publications

The Alteration of Subtelomeric DNA Methylation in Aging-Related Diseases

The Alteration of Subtelomeric DNA Methylation in Aging-Related Diseases

Association of birth outcomes and postnatal growth with adult leukocyte telomere length: Data from New Delhi Birth Cohort

Telomere Attrition in Neurodegenerative Disorders

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