Primer group for detecting BMPR1B gene mutation, purpose of primer group and kit containing primer group

A technology of primer set and kit, which is applied in the direction of recombinant DNA technology, microbial measurement/testing, biochemical equipment and methods, etc., can solve problems such as unreachable detection, ensure accuracy and uniqueness, and have a wide range of applications , the effect of improving the quality of the population

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 BMPR1B 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 BMPR1B gene mutation, purpose of primer group and kit containing primer group
  • Primer group for detecting BMPR1B gene mutation, purpose of primer group and kit containing primer group
  • Primer group for detecting BMPR1B gene mutation, purpose of primer group and kit containing primer group

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] According to the 1st to 10th exon region, 3'UTR, 5'UTR and promoter sequence of the BMPR1B gene published by NCBI Gene Bank (NG_009245), a primer set for detecting whether the BMPR1B gene is mutated was designed. The set includes a PCR amplification primer set and a PCR sequencing primer set corresponding to the above regions, and the results are shown in Table 1 and Table 2.

Embodiment 2

[0032] A kit for detecting BMPR1B gene mutations, including PCR amplification primer sets and PCR sequencing primer sets corresponding to the 1st to 10th exon regions, 3'UTR, 5'UTR and promoter sequences of the BMPR1B gene, a total of 24 each For the primers 1OD, refer to Table 1 and Table 2 for the sequences of each pair of primers.

[0033] This kit is stored at -20°C, and repeated freezing and thawing should be avoided as much as possible.

Embodiment 3

[0035] Kits for detecting BMPR1B gene mutations, including:

[0036] (1) PCR amplification primer set and PCR sequencing primer set for the 1st to 10th exon region, 3'UTR, 5'UTR and promoter sequence of BMPR1B gene, a total of 24 pairs, each primer 1OD, each pair of primer sequences See Tables 1 and 2.

[0037] (2) PCR amplification reagent: 2.5mM dNTP mixture 40μl, 5×amplification buffer (containing Mg 2+ ) 100μl, 5Units / ul Taq DNA polymerase 5μl.

[0038] (3) PCR product purification reagents: 1 U / ul SAP enzyme 20 μl, 10 U / ul ExoI enzyme 10 μl.

[0039] (4) PCR sequencing reagents: BigDye 3.1 mix 50μl, 0.125M EDTA solution 50μl, absolute ethanol 1ml, 75% ethanol solution 1.5mL, HIDI solution 500μl.

[0040] 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 BMPR1B gene mutation. The primer group comprises a BMPR1B gene first to tenth exon regions, a PCR (polymerase chain reaction) amplification primer group corresponding to 3'UTR, 5'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 BMPR1B gene mutation occurs or not; the accuracy and the uniqueness of the detection result are ensured.

Description

technical field [0001] The invention relates to primers, in particular to a primer set for detecting BMPR1B gene mutation, and also relates to the use of the primer set and a kit containing the primer set. Background technique [0002] BMPR1B (bone morphogenetic protein receptor, type IB) is a member of the transforming growth factor β (TGF-β) receptor superfamily and is a transmembrane serine / threonine kinase receptor. The signal transduction system of the TGF-β superfamily has a variety of biological functions, including the regulation of cell function and tissue cell differentiation. [0003] Pulmonary hypertension is a group of clinicopathophysiological syndromes caused by various reasons and the mean pulmonary artery pressure (mPAP) measured by right heart catheterization in resting state (meanpulmonary arterial pressure, mPAP) ≥ 25mmHg. Pulmonary hypertension can exist independently as a disease, and more commonly it is a pathophysiological manifestation of many disea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
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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