Method for accurately measuring length of crowd telomere

A technology for accurate measurement of telomere length, which is applied in biochemical equipment and methods, microbiological determination/inspection, etc. It can solve the problems of large sample variability, difficult comparison of telomere length, large error, etc., and achieve DNA consumption The effect of less and faster measurement

Inactive Publication Date: 2017-05-31
上海三誉华夏基因科技有限公司
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Problems solved by technology

The main limitation of RT-qPCR is that the variability between samples is large, and the error in the pipetting process is large. In addition, this method also needs to make assumptions, inclu

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  • Method for accurately measuring length of crowd telomere
  • Method for accurately measuring length of crowd telomere
  • Method for accurately measuring length of crowd telomere

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Embodiment Construction

[0020] The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative work shall fall within the protection scope of the present invention.

[0021] 1. Extraction of human genomic DNA

[0022] 150-200μl of blood was collected by venipuncture, and genomic DNA of human samples was extracted with Beijing Tiangen Blood Genome Extraction Kit.

[0023] Take 5μl of cell DNA extraction solution, add 495μl of tri-distilled water to dilute 500 times, and measure the OD value of 260nm and 280nm wavelength by UV spectrophotometer. The calculation formula of DNA concentration is as follows:

[0024] The concentration of DN...

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Abstract

The invention relates to the technical field of molecular organism and specifically relates to a method for accurately measuring length of crowd telomere. A 36B4 gene of a coding acidic ribosome phosphoprotein is selected as a single copy gene. The method comprises the following steps: extracting a human genome DNA; designing a reference substance and a primer and compounding; designing telomere and 36B4 standard curve; adopting a RT-qPCR method for measuring the length of crowd telomere; analyzing data. According to the invention, on the basis of the RT-qPCR method, the telomere standard curve and the single copy gene standard curve are added for more accurately measuring T and S values, so that a more accurate telomere length can be acquired; the DNA dosage is small; only 60ng DNA is required; the measuring speed is high; the measurement has comparability; the telomere length can be accurately measured in batches; and the method is specially suitable for the measurement for the crowd telomere length.

Description

Technical field [0001] The invention relates to the field of molecular biology technology, and specifically is a method for accurately measuring the length of human telomeres. Background technique [0002] Telomeres, a special nuclear protein complex, are composed of the evolutionarily conserved tandem repeat DNA sequence TTAGGG, located at the end of eukaryotic chromosomes, and play an important role in eukaryotic cell biology. The shortening of telomeres acts as a mitotic counter and life-limiting function, and can reflect organ aging at the cellular level. The study of telomere length is extremely important for the research of telomere and telomere-related diseases and even tumors. Telomere length measurement is an important method to study the biology of telomeres and the influence of telomere dysfunction on degenerative diseases. Clinically, telomere length measurement is widely used to diagnose telomere-related diseases, including cardiovascular disease, progressive liver...

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/686C12Q2521/107
Inventor 薛京伦汪旭王晗杨国防马晓玲
Owner 上海三誉华夏基因科技有限公司
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