1. High-throughput single telomere analysis using DNA microarray and fluorescent in situ hybridization.
- Author
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Zheng, Yun-Ling, Wu, Xingjia, Williams, Madeline, Verhulst, Simon, Lin, Jue, Takahashi, Yusuke, Ma, Jian-Xing, and Wang, Ying
- Subjects
In Situ Hybridization ,Fluorescence ,Humans ,Telomere ,Oligonucleotide Array Sequence Analysis ,Telomere Homeostasis ,High-Throughput Screening Assays ,Male ,Adult ,Female ,Middle Aged - Abstract
The human telomere system is highly dynamic. Both short and long leucocyte average telomere lengths (aTL) are associated with an increased risk of cancer and early death, illustrating the complex relationship between TL and human health and the importance of assessing TL distributions with single TL analysis. A DNA microarray and telomere fluorescent in situ hybridization (DNA-array-FISH) approach was developed to measure the base-pair (bp) lengths of single telomeres. On average 32000 telomeres were measured per DNA sample with one microarray chip assaying 96 test DNA samples. Various telomere parameters, i.e. aTL and the frequency of short/long telomeres, were computed to delineate TL distribution. The intra-assay and inter-assay coefficient of variations of aTL ranged from 1.37% to 3.98%. The correlation coefficient (r) of aTL in repeated measurements ranged from 0.91 to 1.00, demonstrating high measurement precision. aTLs measured by DNA-array-FISH predicted aTLs measured by terminal restriction fragment (TRF) analysis with r ranging 0.87-0.99. A new accurate and high-throughput method has been developed to measure the bp lengths of single telomeres. The large number of single TL data provides an opportunity for an in-depth analysis of telomere dynamics and the complex relationship between telomere and age-related diseases.
- Published
- 2024