Primer group for detecting ALK-1 (activin receptor type II-like kinase-1) gene mutation, and application thereof as well as kit comprising primer group

A primer set and kit technology, applied in the direction of DNA/RNA fragment, recombinant DNA technology, microbial determination/inspection, etc., can solve dangerous problems and achieve the effect of ensuring accuracy and uniqueness, and the method is simple

Active Publication Date: 2013-03-27
广州瑞能精准医学科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] At present, patients with pulmonary arterial hypertension with ALK1 gene mutation are younger than patients with bone morphogenic protein 2 (BMPR2) gene mutation, and once the disease progresses more rapidly, it is more dangerous

Method used

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  • Primer group for detecting ALK-1 (activin receptor type II-like kinase-1) gene mutation, and application thereof as well as kit comprising primer group
  • Primer group for detecting ALK-1 (activin receptor type II-like kinase-1) gene mutation, and application thereof as well as kit comprising primer group
  • Primer group for detecting ALK-1 (activin receptor type II-like kinase-1) gene mutation, and application thereof as well as kit comprising primer group

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Embodiment 1

[0046] According to the 1~10 exon region, 3'UTR and promoter sequence of ALK-1 published by NCBI Gene Bank (NC-000012), a primer set for detecting whether the ALK-1 gene is mutated was designed. The set includes the PCR amplification primer set and the PCR sequencing primer set of the above regions, and the results are shown in Table 1 and Table 2.

Embodiment 2

[0048] A kit for detecting ALK-1 gene mutation, comprising the following components:

[0049] There are 15 pairs of PCR amplification primers and PCR sequencing primers corresponding to the 1st to 10th exon regions, 3'UTR and promoter sequences of the ALK-1 gene, and each primer is 1OD. For the primer sequences of each pair, see Table 1 and Table 2.

[0050] This kit is stored at -20°C, and repeated freezing and thawing should be avoided as much as possible.

Embodiment 3

[0052] A kit for detecting ALK-1 gene mutation, comprising the following components:

[0053] (1) PCR amplification primer set and PCR sequencing primer set corresponding to the 1st to 10th exon region, 3'UTR and promoter sequence of ALK-1 gene, a total of 15 pairs, each primer 1OD, each pair of primers See Table 1 and Table 2 for the sequence.

[0054] (2) PCR amplification reagent: 40 μl of 2.5mM dNTP mixture, 5× amplification buffer (containing Mg 2+ ) 100 μl, 5 Units / ul Taq DNA polymerase 5 μl.

[0055] (3) PCR product purification reagents: 1U / ul SAP enzyme 20 μl, 10 U / ul ExoI enzyme 10 μl.

[0056] (4) PCR sequencing reagents: BigDye3.1mix 50μl, EDTA solution (0.125M) 50μl, absolute ethanol 1ml, ethanol solution (75%) 1.5mL, HIDI solution 500μl.

[0057] 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 ALK-1 (activin receptor type II-like kinase-1) gene mutation, at least comprising ALK-1 gene first to tenth exon regions, and a PCR (Polymerase Chain Reaction) amplification primer group and a PCR sequencing primer group which correspond to any region of a 3' UTR (Untranslated Region) and a promoter. The invention further discloses an application of the primer group, a kit comprising the primer group, and a method for carrying out PCR amplification on an ALK-1 gene by the primer group. The primer group can be used for specially identifying whether the ALK-1 gene is mutated or not so as to ensure the accuracy and the uniqueness of a detected result.

Description

technical field [0001] The invention relates to primers, in particular to a primer set for detecting ALK-1 gene mutation, and also relates to the use of the primer set and a kit containing the primer set. Background technique [0002] Activin receptor kinase 1 (activin receptor type II-like kinase-1, ALK-1, also known as ACVRL1) is located at 12q11-q14, contains 10 exons, and the genome length is about 15kb+. Its mutations can cause hereditary hemorrhagic telangiectasia (HHT) or hereditary pulmonary arterial hypertension (heritable pulmonary arterial hypertension PAH). Some mutation carriers have both conditions at the same time. HHT is an autosomal dominant genetic hemorrhagic disorder characterized by multiple mucous telangiectasia with recurrent hemorrhage. Clinically, it can be generally divided into two phenotypes: type I HHT and type II HHT. Among them, type II HHT is caused by ALK-1 gene mutation. The most common clinical manifestation of HHT-II is repeated spontane...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
Inventor 卢文菊张晨婷王健
Owner 广州瑞能精准医学科技有限公司
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