Relative Human Telomere Length Quantification by Real-Time PCR

Vasilishina, A.; Kropotov, A.; Spivak, I.; Bernadotte, A.

Авторы: Vasilishina A., Kropotov A., Spivak I., Bernadotte A.
Сборник: Cellular Senescence: Methods and Protocols, Methods in Molecular Biology
Том: 1896
Глава в коллективной монографии
Год издания: 2019
Место издания: Springer Nature 11 WEST 42ND STREET, NEW YORK, USA, NY,10036
Первая страница: 39
Последняя страница: 44
DOI: 10.1007/978-1-4939-8931-7_5
Аннотация: Telomere measurement by quantitative PCR amplification is a well-known simple method to detect telomere length that involves large numbers of samples. The method has been developed by Cawthon in 2002 (Cawthon, Nucleic Acids Res 30:47e–47, 2002) and remains the most frequently used technique either in original or modified version. Telomere length is estimated by comparing the amount of telomere repeat amplification product (T) to a single copy gene (S) product. The T/S ratio correlates with the average telomere length. Cawthon suggested and recommended the use of 36B4 (RPLP0) as a single copy gene. However, Cawthon’s suggestion was no longer considered a single copy gene and the gene was not suitable and appropriate for normalization. We thereby introduced a simple method for relative measurement of average human telomere length using quantitative real-time PCR. Our protocol was based on Cawthon’s initial technique (Cawthon, Nucleic Acids Res 30:47e–47, 2002), modified by single-copy gene (SCG) primers and optimized. This technique is rapid, low cost, not demanding on DNA amount (or live cells), and can be used for a high-throughput screening and time monitoring. Key words Telomere length, Quantitative PCR, Real-time PCR, Telomere, Telomere measurement, IFNB1, 36B4, Primers, Single-copy gene
Добавил в систему: Бернадотт Александра

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Relative Human Telomere Length Quantification by Real-Time PCRстатья

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