Method and kit for detecting ADRB1 gene single nucleotide polymorphic site

A technology of polymorphic sites and single nucleotides, applied in the field of molecular biology, to achieve simple and objective interpretation of results, strong specificity, and authentic and reliable results

Inactive Publication Date: 2017-06-13
重庆迪威纳生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Beta-adrenoceptor blockers (beta-blockers), such as metoprolol and bucinolol, have been shown to increase cardiac The survival rate of patients with vascular diseases and the reduction of mortality are the most commonly used first-line drugs for the clinical treatment of hypertension, but the therapeutic effect is affected by the polymorphism of the β1-adrenergic receptor (ADRB1) heterogeneity in

Method used

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  • Method and kit for detecting ADRB1 gene single nucleotide polymorphic site
  • Method and kit for detecting ADRB1 gene single nucleotide polymorphic site
  • Method and kit for detecting ADRB1 gene single nucleotide polymorphic site

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Experimental program
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Effect test

Embodiment 1

[0052] Embodiment 1 primer and probe synthesis

[0053] Design and synthesize amplification primers for ADRB1rs1801252 and ADRB1rs1801253, 145AARMS-specific amplification primers and specific probes for ADRB1rs1801252; 145G ARMS-specific amplification primers and specific probes for ADRB1rs1801252; ADRB1rs1801253 1165G ARMS-specific amplification primers and specific probes for the site; 1165CARMS-specific amplification primers and specific probes for the ADRB1rs1801253 site. (The reference gene sequence for designing primers is derived from NCBI database).

[0054] The nucleotide sequence of the upstream amplification primer of the ADRB1rs1801252 site is shown in SEQ ID NO: 1, the nucleotide sequence of the downstream amplification primer is shown in SEQ ID NO: 2; the nucleotide sequence of the 145AARMS specific amplification primer The sequence is shown in SEQ ID NO: 3, the nucleotide sequence of the specific probe is shown in SEQ ID NO: 4; the nucleotide sequence of the 14...

Embodiment 2

[0058] Embodiment 2 prepares internal standard system

[0059] Design and synthesize a pair of internal standard primers designed for the human genome, the internal standard forward primer is shown in SEQ ID NO:13, the internal standard reverse primer is shown in SEQ ID NO:14; design and synthesize the internal standard probe , the probe is SEQ ID NO:15. The internal standard primers were prepared into 100uM mother solution for storage, and the internal standard probe was prepared into 100uM mother solution for storage.

[0060] No. Nucleotide sequence SEQ ID NO: 13 5'-CGCGAACTCACCGT-3' SEQ ID NO: 14 5'-CACTAGGCGCTCACTGT-3' SEQ ID NO: 15 5'-ROX-CACCTTCCCCATGGTGTCT-BHQ2-3'

Embodiment 3

[0061] Embodiment 3 prepares other reagents

[0062] Prepare PCR buffer containing 1.0 mM MgCl 2 , dATP, dUTP, dCTP and dGTP each 1.0mM; prepare enzyme mixture, which contains Taq enzyme 0.5×10 3 U / ml, UNG enzyme 0.1×10 3 U / ml.

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Abstract

The invention belongs to the technical field of molecular biology, and particularly relates to a method for detecting an ADRB1 gene single nucleotide polymorphic site. The method comprises the following specific steps: 1) design of a specific primer: designing a specific amplification primer, an ARMS specific amplification primer, an interior label amplification primer, a specific probe and an interior label probe according to a proved ADRB1 gene label single nucleotide polymorphic site, wherein the 5' end of the specific probe carries a fluorescent group, and the 3' end of the specific probe carries a fluorescent quenching group; 2) acquiring DNA (Deoxyribonucleic Acid) of a sample genome to be detected; 3) fluorescent quantitative PCR (Polymerase Chain Reaction) amplification: performing a PCR reaction by taking the DNA of a sample to be detected as a template with the specific amplification primer in the step 1), and analyzing a detection result. The method has high sensitivity and high specificity, and is easy and convenient to operate; detection can be finished within 90 minutes, and a result interpretation method is simple, objective and convenient for analysis.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to a method and a kit for detecting single nucleotide polymorphism sites of ADRB1 gene. Background technique [0002] Cardiovascular diseases such as high blood pressure seriously threaten human health. They are common diseases among middle-aged and elderly people over 50 years old, and have the characteristics of high prevalence, high disability rate and high mortality rate. According to conservative estimates, the number of hypertensive patients in my country exceeds 160 million, and the incidence rate is increasing year by year. Treatment of hypertension and its complications has become the third largest economic burden worldwide. Drugs are the main means of treating hypertension and its complications at present and for a long period of time in the future. [0003] However, due to the physiological state of the patient (such as age, gender, race, obesity, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6851C12Q1/6858C12Q1/6883C12Q2600/106C12Q2600/156C12Q2600/16C12Q2535/137C12Q2561/101C12Q2531/113C12Q2537/143C12Q2545/101
Inventor 不公告发明人
Owner 重庆迪威纳生物技术有限公司
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