Telomere length detection primer composition and kit and application thereof

A technology of primer composition and detection end, which is applied in the field of telomere length detection primer composition, can solve the problems of poor accuracy and repeatability, and achieve the effect of preventing mutual fusion and ensuring complete replication

Inactive Publication Date: 2022-01-21
中科解码北京生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the accuracy and repeatability of the above methods are poor

Method used

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  • Telomere length detection primer composition and kit and application thereof
  • Telomere length detection primer composition and kit and application thereof
  • Telomere length detection primer composition and kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1. A primer composition for detecting telomere length

[0051] Selection of internal reference genes: Interferon-encoding gene IFNB1, eukaryotic translation elongation factor EEF1G, peroxide oxidoreductase PRDX1, ribosomal protein RPSA and / or cytoskeleton protein VIM were selected as internal reference genes.

[0052] The primer composition is used to detect telomeric repeat sequence Tel, interferon coding gene IFNB1, eukaryotic translation elongation factor EEF1G, peroxide oxidoreductase PRDX1, ribosomal protein RPSA and / or cytoskeleton protein VIM.

[0053] (1) The primer composition of the telomeric repeat sequence Tel is:

[0054] F (SEQ ID NO: 1): 5'-CGGTTTGTTTGGGTTTGGGTTTGGGTTTGGGTTTGGGTT-3';

[0055] R (SEQ ID NO:2): 5'-GGCTTGCCTTACCCTTACCCTTACCCTTACCCTTACCCT-3';

[0056] (2) The primer composition of the interferon-encoding gene IFNB1 is:

[0057] F (SEQ ID NO:3): 5'-ATGACCAACAAGTGTCTCCTCC-3';

[0058] R (SEQ ID NO:4): 5'-GGAATCCAAGCAAGTTGTAGCTC-3'; ...

Embodiment 2

[0074] Example 2. A kit for detecting telomere length

[0075] (1) Comprising the primer composition described in Example 1.

[0076] (2) Also includes PCR enzyme, buffer, dNTP, template and deionized water, etc.

[0077] Among them, the 10μL reaction system is as follows: 2×ChamQ SYBR qPCR Master Mix, 5μL; 50×ROXReference Dye, 0.2μL; Genomic DNA, 20-40ng; Telomere repeat amplification primer 10μM, 0.2μL; μL.

Embodiment 3

[0078] Example 3. A method for detecting telomere length

[0079] 1. Extract saliva / throat swab / blood genomic DNA

[0080] Taking saliva samples as an example, genomic DNA was extracted using a buccal swab genomic DNA extraction kit (Tiangen Biochemical Technology Co., Ltd., DP322), and the operation was performed strictly according to the instructions.

[0081] 1.1. Processing materials: Take 200-400 μL of saliva sample according to the requirements, add an equal volume of buffer GHA, and mix well by inversion. Then take out 300-350 μL of the sample, add 10 μL of Proteinase K solution, vortex for 10 seconds to mix, place at 65°C for 30 minutes, and vortex every 10 minutes to mix.

[0082] 1.2. Add 500 μL buffer GBS, mix thoroughly by inversion, and place at room temperature for 5 minutes.

[0083] 1.3. Add the solution obtained in the previous step into an adsorption column CB2, centrifuge at 12000rpm for 30s, discard the waste liquid in the collection tube, and put the ads...

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Abstract

The invention belongs to the technical field of biological detection, and particularly relates to a telomere length detection primer composition and a kit and application thereof. The kit provided by the invention comprises a primer composition for detecting a telomere repetitive sequence Tel, an interferon coding gene IFNB1, an eukaryotic translation elongation factor EEF1G, a peroxide oxidoreductase PRDX1, a ribosomal protein RPSA and / or a cytoskeleton protein VIM. The primer composition provided by the invention is used for detecting the telomere length, has better sensitivity, accuracy and repeatability, is high in amplification efficiency, simple and convenient to operate, rapid and time-saving, can be used for high-throughput screening and time monitoring, and has important practical significance for genetic and biological age evaluation of cancers and molecular epidemiological research.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to a primer composition for detecting telomere length, a kit and application thereof. Background technique [0002] The theory of telomeres was proposed by Alexei Olovnikov in 1971. Telomeres are DNA-protein complexes that exist at the end of chromosomes in eukaryotic cells. Their main function is to maintain the integrity and stability of chromosomes. Telomeres are composed of short double-stranded repeat sequences TTAGGG in series, and the length of the double-stranded region is generally 0.5-20kb. Telomeres protect chromosomes from being degraded by nucleases, prevent chromosomes from merging with each other, and solve the terminal inversion of DNA replication to ensure the complete replication of chromosomes. Without telomere protection, chromosomes would gradually lose genetic information as cells divide and replicate. Telomeres are the molecular cloc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12Q1/6886C12Q1/6888C12Q1/6851C12N15/11
CPCC12Q1/6883C12Q1/6886C12Q1/6888C12Q1/6851C12Q2600/156C12Q2600/166C12Q2531/113C12Q2563/107C12Q2525/151C12Q2545/101
Inventor 段炼
Owner 中科解码北京生物技术有限公司
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