Specific primer group, test kit and test method for testing polymorphic sites of human MTHFR and MTRR genes

A gene polymorphism, human detection technology, applied in biochemical equipment and methods, recombinant DNA technology, microbial determination/inspection, etc. It can reduce the pain, use less blood, and reduce economic expenses.

Inactive Publication Date: 2018-12-21
XIAMEN WIZ BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

These methods either have limited detection capabilities, or are time-consuming and labor-intensive. More importantly, on the one hand, these methods cannot simultaneously perform high-throughput detection of multiple

Method used

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  • Specific primer group, test kit and test method for testing polymorphic sites of human MTHFR and MTRR genes
  • Specific primer group, test kit and test method for testing polymorphic sites of human MTHFR and MTRR genes
  • Specific primer group, test kit and test method for testing polymorphic sites of human MTHFR and MTRR genes

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Experimental program
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Embodiment 1

[0039] Embodiment 1: the preparation of kit

[0040] 1. Design and synthesis of primers and probes

[0041] For the MTHFR (C677T), MTRR (A66G), and MTHFR (A1298C) gene loci in the human genome (for the sequence, refer to the human genome sequence published in the NCBI database), specific primers and probes were designed using Primer Premier 3.0 software. Specific primers and probe sequences are shown in Table 1:

[0042] Table 1:

[0043]

[0044] 2. Selection of reference substance

[0045]The wild-type control is the wild-type plasmid mixture of the three gene loci of MTHFR (C677T), MTHFR (A1298C), and MTRR (A66G); the negative control is 1*TE buffer or nuclease-free water.

[0046] 3. Composition of PCR reaction solution

[0047] The PCR reaction solution includes the above-mentioned specific primers and probes, wherein:

[0048] The lyophilized powder in the reaction tube contains M677 forward primer F1, M677 reverse primer R1, M677 probe P1, M1298 forward primer F...

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Abstract

The invention discloses a specific primer group for testing polymorphic sites of human MTHFR and MTRR genes, and the primer group comprises a specific primer and a probe for the sites of MTHFR (C677T), MTHFR (A1298C) and MTRR (A66G) genes; and the invention also relates to a kit for testing the polymorphic sites of human MTHFR and MTRR genes, the kit comprises lyophilized powder (10 x SSP buffer,dNTPs, MgCl2, Taq enzyme and trehalose) of PCR reaction liquid containing the primer and the probe, and the kit is stored at 2-30 DEG C away from light. The invention also relates to a test method fortesting the polymorphic sites of human MTHFR and MTRR genes, and the method avoids a procedure of DNA extraction, and directly adopts a microamount of whole blood for amplification, which requires alittle blood and reduces the pain of patients; and the method saves time and effort, and greatly reduces the economic expenditure.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a specific primer set, a kit and a detection method for detecting human MTHFR and MTRR gene polymorphism sites. Background technique [0002] Folic acid is a water-soluble B vitamin composed of pteridine, p-aminobenzoic acid and glutamic acid residues. It is a substance necessary for the growth and reproduction of body cells. It participates in the synthesis of other important compounds such as DNA, RNA, and protein. , play an important role in the body's metabolic process. Humans such as lack of folic acid can cause megaloblastic anemia and leukopenia, can cause hyperhomocysteinemia and other symptoms. Folic acid is especially important for pregnant women. The demand for folic acid in pregnant women is 4 times higher than that of normal people. If there is a metabolic disorder of folic acid in pregnant women and pregnant women, it can lead to open neural tube defects (such as anenc...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/156
Inventor 林志铿周晓光李运涛杨璐平雷洋王松林
Owner XIAMEN WIZ BIOTECH CO LTD
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