The Method of Measuring the Absolute Length of Chromosome Telomeres by Single-color Multiplex Quantitative PCR

Abstract

The invention discloses a method for measuring the absolute length of chromosomal telomeres by single-color multiplex quantitative PCR, which is characterized in that the method comprises the following steps: (1) designing and screening primers for chromosomal telomeres and primers for amplifying internal reference genes Single-copy gene primers require that the difference between the Tm values ​​of the telomere gene primers and the single-copy gene primers be in the range of 10-20°C; (2) Take the telomere sequence of SEQNo: 1 and the single-copy gene of SEQNo: 2 as Standard sequence, draw a standard curve by the external standard method; (3) extract the template DNA, add the template DNA to the PCR reaction system with the primers in step (1) to perform PCR amplification of the target gene in the template DNA; (4) use the standard The curve is the control. The absolute length of telomere was determined by quantitative PCR method. This method eliminates the sampling error, makes the results more accurate, and breaks through the limitation that ordinary quantitative PCR can only measure the relative length of telomeres.

Description

technical field [0001] The invention belongs to the technical field of gene measurement, and in particular relates to a method for measuring the absolute length of chromosomal telomeres by a single-color multiple quantitative PCR method. Background technique [0002] Telomeres (tolemere) are DNA repeats at the ends of chromosomes, which are tandem repeats of (TTAGGG) in human chromosomes. Telomeres are specialized structures at the ends of chromosomes that, in normal human cells, gradually shorten as cells divide. The role of telomeres is to maintain the integrity of chromosomes. Once the cell divides, because the direction of DNA replication must be from 5' to 3', the telomere shortens a little each time the DNA replicates. Once telomeres are depleted, chromosomes are prone to mutations that lead to arteriosclerosis and certain cancers. In recent years, a variety of telomere length measurement methods have been reported, including Southern blot (SB), hybridization protec...

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Problems solved by technology

[0004] The purpose of the present invention is to provide a method for measuring the absolute length of chromosomal telomeres by a single-color multiplex quantitative PCR method, which solves the problems of time-consuming and labor-consuming measurement of the absolute length of chromosomal telomeres, the need for a large number of DNA samples, and low measurement accuracy in the prior art.

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