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Primer group for detecting CAV1 gene mutation, purpose of primer group and kit containing primer group

A primer set and kit technology, applied in the field of primer sets, can solve problems such as failure to achieve detection, and achieve the effects of ensuring accuracy and uniqueness, improving population quality, and widening detection range.

Inactive Publication Date: 2016-08-24
THE FIRST AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV (GUANGZHOU RESPIRATORY CENT)
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The position and mode of CAV1 gene mutation are different for different people, and the purpose of detection cannot be achieved by fluorescent quantitative PCR method

Method used

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  • Primer group for detecting CAV1 gene mutation, purpose of primer group and kit containing primer group
  • Primer group for detecting CAV1 gene mutation, purpose of primer group and kit containing primer group
  • Primer group for detecting CAV1 gene mutation, purpose of primer group and kit containing primer group

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Experimental program
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Effect test

Embodiment 1

[0028] According to the 1~3 exon region, 3'UTR and promoter sequence of CAV1 disclosed by NCBI Gene Bank (NC-000012), a primer set for detecting whether the CAV1 gene is mutated is designed, and the primer set includes the above The PCR amplification primer set and PCR sequencing primer set of the region, the results are shown in Table 1 and Table 2.

Embodiment 2

[0030] A kit for detecting CAV1 gene mutation, comprising the following components:

[0031] There are 11 pairs of PCR amplification primers and PCR sequencing primers corresponding to the 1st to 3rd exon regions, 3'UTR and promoter sequences of the CAV1 gene, and each primer is 1OD. For the sequences of each pair of primers, see Table 1 and Table 2.

[0032] This kit is stored at -20°C, and repeated freezing and thawing should be avoided as much as possible.

Embodiment 3

[0034] A kit for detecting CAV1 gene mutation, comprising the following components:

[0035] (1) There are 11 pairs of PCR amplification primers and PCR sequencing primers corresponding to the 1st to 3rd exon regions, 3'UTR and promoter sequences of the CAV1 gene, and each primer is 1OD. For the primer sequences of each pair, see Table 1 and Table 2.

[0036] (2) PCR amplification reagent: 2.5mM dNTP mixture 40μl, 5×amplification buffer (containing Mg 2+ ) 100μl, 5Units / ul Taq DNA polymerase 5μl.

[0037] (3) PCR product purification reagents: 1 U / ul SAP enzyme 20 μl, 10 U / ul ExoI enzyme 10 μl.

[0038] (4) PCR sequencing reagents: BigDye 3.1 mix 50μl, EDTA solution (0.125M) 50μl, absolute ethanol 1ml, ethanol solution (75%) 1.5mL, HIDI solution 500μl.

[0039] This kit is stored at -20°C, and repeated freezing and thawing should be avoided as much as possible.

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Abstract

The invention discloses a primer group for detecting CAV1 gene mutation. The primer group comprises a CAV1 gene first to third exon regions, a PCR (polymerase chain reaction) amplification primer group corresponding to 3'UTR and a starter and a PCR sequence testing primer. The invention also discloses a purpose of the primer group and a kit containing the primer group. The primer group can be used for specifically identifying whether the CAV1 gene mutation occurs or not; the accuracy and the uniqueness of the detection result are ensured.

Description

technical field [0001] The invention relates to a primer set, in particular to a primer set for detecting CAV1 gene mutation. The present invention also relates to the use of the primer set and a kit comprising the primer set. Background technique [0002] CAV1 (caveolae protein), a transmembrane protein of 21kDa to 24kDa, participates in the formation of cell membrane structure caveolae, and its 82-101 amino acid residue region is called Caveolin-scaffolding Domain (SCD), which is the structural basis for interaction with various other molecules. CAV1 is widely distributed in the body's terminally differentiated cells or quiescent cells. Not only can it participate in the regulation of signal transduction, inhibit cell growth and transformation, but also participate in vesicle transport (including endocytosis, pinocytosis and transcellular action) and maintenance of cholesterol homeostasis. Its mutation can cause hereditary pulmonary arterial hypertension (heritablepulmon...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6883C12Q2600/156
Inventor 卢文菊张晨婷王健
Owner THE FIRST AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV (GUANGZHOU RESPIRATORY CENT)
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