Primer probe group for detecting rs1045642 and application of primer probe group

A primer-probe and probe technology, applied in recombinant DNA technology, microbial assay/inspection, biochemical equipment and methods, etc., can solve problems such as resistance and drug concentration reduction, and achieve simple operation and great application and promotion value. Effect

Inactive Publication Date: 2018-05-22
山东德诺生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Individuals carrying mutant alleles have changes in the activity of P-glycoprotein, resulting in changes in the transmembrane transport process of various drugs mediated by it in vivo, which makes t

Method used

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  • Primer probe group for detecting rs1045642 and application of primer probe group
  • Primer probe group for detecting rs1045642 and application of primer probe group
  • Primer probe group for detecting rs1045642 and application of primer probe group

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Embodiment 1, be used to detect the design of the primer probe set of rs1045642

[0051] After a lot of design, pre-experiment, and effect comparison, a set of primer probes for detecting rs1045642 was obtained.

[0052] Primer F: 5'-TTGCTGAGAACATTGCCTATGGAGA-3';

[0053] Primer R: 5'-TGACTCGATGAAGGCATGTATGTTG-3';

[0054] Probe P: 5'-FAM- ATC GTGAGGGCAGCAAAG GAT -BHQ1-3'.

[0055] Primer F is a single-stranded DNA molecule shown in Sequence 1 of the Sequence Listing. Primer R is a single-stranded DNA molecule shown in Sequence 2 of the Sequence Listing. Probe P is a single-stranded DNA molecule with a FAM fluorescent group at the 5' end and a BHQ1 fluorescent quencher group at the 3' end. The nucleotide sequence of the DNA molecule is shown in sequence 3 in the sequence table, and the underlined nucleosides The acids (ie, positions 1, 2, 3, 19, 20, and 21) are locked nucleic acids (LNA).

[0056] The target sequences of primer F and primer R are shown in seque...

Embodiment 2

[0057] Embodiment 2, the application of the primer probe set for detecting rs1045642

[0058] 1. Blood sample processing (processing method of a single blood sample)

[0059] 1. Take 65 μL of EDTA anticoagulant blood, add 500 μL of lysate, mix by inversion, centrifuge at 12000 rpm for 4 min, and discard the supernatant.

[0060] Lysis solution: 1.7g / 100ml NaCl aqueous solution.

[0061] 2. After completing step 1, add 120 μL of preservation solution, first bathe in water at 65°C for 10 minutes, then in water bath at 85°C for 10 minutes, then centrifuge at 12,000 rpm for 4 minutes, then store at 4°C for later use, and take the supernatant before use, which is the template sample.

[0062] The preparation method of the preservation solution: ① mix 25 parts by volume of FG buffer solution and 4 parts by volume of proteinase K solution to obtain a mixed solution; ② mix 1 part by volume of the mixed solution obtained in step ① with 5 parts by volume of ddH 2 O mixed to obtain a p...

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Abstract

The invention discloses a primer probe group for detecting rs1045642 and application of the primer probe group. The invention firstly protects the primer probe group which comprises a primer F, a primer R and a probe P; the primer F is a single-strand DNA molecule shown as a sequence 1 in a sequence table; the primer R is a single-strand DNA molecule shown as a sequence 2 in the sequence table; the probe P is a single-strand DNA molecule with the one terminus having a fluorescent group and the other terminus having a fluorescence quenching group; a part of nucleotides in the DNA molecule are locked nucleic acids; and a nucleotide sequence of the DNA molecule is shown as a sequence 3 in the sequence table. The primer probe group provided by the invention can be used for detecting rs1045642and judging a genotype of a sample to be detected based on the site so as to guide medication and having an important application and popularization value.

Description

technical field [0001] The invention relates to a primer probe set for detecting rs1045642 and application thereof. Background technique [0002] ATP-binding cassette transporters (ABC transporters) transport a variety of molecules to and from the cell membrane and are divided into seven distinct subfamilies (ABC1, MDR / TAP, MRP, ALD, OABP, GCN20, White). Members of the MDR / TAP family are involved in the multidrug resistance process. P-glycoprotein (P-gp) is a member of the MDR / TAP family. P-glycoprotein is an ATP-dependent drug efflux pump responsible for reducing drug accumulation in drug-resistant cells and mediating the formation of drug resistance to anticancer drugs. P-glycoprotein plays an important role in the absorption process of drugs. P-glycoprotein on the surface of intestinal mucosa affects drug absorption and indirectly affects blood drug concentration by pumping drugs back into the intestinal cavity. P-glycoprotein also acts as a transporter in the blood-br...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12Q1/6883C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/6883C12Q1/6886C12Q2600/156C12Q2563/107C12Q2545/114C12Q2527/107C12Q2525/10
Inventor 丛茜赵海文刘婷婷李朋孙文文杨银辉
Owner 山东德诺生物科技有限公司
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