Method for measuring telomere absolute length

A technology for measuring telomere length and measuring ends, which is applied in the field of life sciences to achieve the effects of predicting disease development and aging, strong repeatability, and less DNA substrates

Inactive Publication Date: 2013-08-07
南京优而生物科技发展有限公司
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

There is no report on the measurement method of the absolute length of telomere

Method used

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  • Method for measuring telomere absolute length
  • Method for measuring telomere absolute length
  • Method for measuring telomere absolute length

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Experimental program
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Effect test

Embodiment

[0033] The embodiment of the present invention will be further described below with reference to the accompanying drawings. This embodiment is aimed at humans with 23 pairs of chromosomes, so the total number of telomeres in the chromosome group is 92.

[0034] 1. Extract genomic DNA from the blood sample, measure the DNA concentration with a Nanodrop spectrophotometer, and store it at 4°C for later use;

[0035] 2. Drawing of telomere length standard curve (such as figure 1 shown):

[0036] The 84mer oligo with 14 repeats 5'-TTAGGG-3' was diluted in a concentration gradient (the concentration gradient was 10 of the defined molar concentration, respectively) -1 to 10 -6 ), the telomere length standard curve was obtained by qPCR reaction, and the PCR primers were as follows:

[0037] tel R: 5'-GGCTTGCCTTACCCTTACCCTTACCCTTACCCTTACCCT-3',

[0038] tel F: 5'-CGGTTTGTTTGGGTTTGGGTTTGGGTTTGGGTTTGGGTT-3'.

[0039]

[0040] In formula (I), c T Indicates the molar concentration...

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Abstract

The invention discloses a method for measuring telomere absolute length. By applying oligomer standard, the telomere absolute length instead of relative length can be measured, which is the largest difference compared with the disclosed real-time quantitative PCR (Polymerase Chain Reaction) method. According to the measuring method, the repeatability is good, few DNA (Deoxyribonucleic Acid) substrates are needed, time is saved, accuracy is realized, and the method is applicable to clinical testing the telomere length and monitoring the dynamic variation, thereby predicting one of disease development and aging indices. The method can be further used for researching preimplantation embryo and stem cell quality.

Description

technical field [0001] The invention relates to the field of life sciences, in particular to a method for measuring the absolute length of telomeres. Background technique [0002] Telomeres are nucleoprotein structures that cap the ends of chromosomes. It is composed of a long hexamer sequence (TTAGGG). The intact telomere structure and its repeating hexamer sequence protect the ends of chromosomes from degradation and play a crucial role in maintaining genome stability. The number of hexameric sequences (TTAGGG) of telomeres decreases with each cell division, resulting in a continuous shortening of telomere length throughout the life of an organism. Shortening of telomere length can lead to fusion of telomere ends and chromosomal instability, thus becoming the cause of various cancers including lung cancer, breast cancer, colon cancer, prostate cancer, leukemia and so on. The shortening of telomere length is also an important factor leading to aging. Studies have shown ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 白丽君金星亮
Owner 南京优而生物科技发展有限公司
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