Quick amplification detection kit of SNP (single nucleotide polymorphism) site of MTRR (5-methyltetrahydrofolate-homocysteinemethyltransferase reductase) and detection method

A technology for detection kits and detection methods, which is applied in the detection of biological samples and in the field of biology, can solve the problems of increasing the economic burden of the testee, high detection cost, and long detection time, so as to reduce economic and mental burden, high detection efficiency, Get the effect in a short time

CN109468371AInactive Publication Date: 2019-03-15XIANGTAN ZHILIAN TECH MATASTASIS PROMOTE CO LTD
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Publication Date
2019-03-15
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention discloses a quick amplification detection kit of an SNP (single nucleotide polymorphism) site of MTRR (5-methyltetrahydrofolate-homocysteinemethyltransferase reductase) and a detection method. The kit comprises an efficient amplification group and a pyrophosphate sequencing group, wherein the efficient amplification group is used for linear exponent PCR (polymerase chain reaction) amplification of the SNP site of MTRR A66G; an amplification product is sequenced by the pyrophosphate sequencing group; a sequence of an amplification primer is shown as SEQ ID NOs (sequence identifiernumbers): 2 and 3 in a sequence table; and a sequence of an annealing primer is shown as SEQ ID NO: 4. A target segment linear exponent amplification method is adopted; the precisely designed amplification primer is high in amplification efficiency and low in mismatching rate; the amplification efficiency is greatly improved; single-stranded DNA (deoxyribonucleic acid) amplified directly can be directly used for pyrophosphate sequencing; the amplification product is not required to be subjected to labeling and double-chain separation; redundant experimental procedures are avoided; and the damage of a strongly basic reagent to an amplification segment is also reduced.
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Description

technical field

[0001] The invention relates to a rapid amplification detection kit and detection method of SNP sites of MTRR, belonging to the field of biotechnology, especially the field of biological sample detection. Background technique

[0002] Methionine synthase reductase MTRR (5-methyltetrahydrofolate-homocysteinemethyltransferase reductase), the full name is encoding 5-methyltetrahydrofolate-homocysteine ​​methyltransferase reductase. Methionine is an essential amino acid for protein synthesis and one-carbon metabolism. Its synthesis is catalyzed by methionine synthase (encoded by the MTR gene), and methionine synthase is eventually oxidized due to the oxidation of the cofactor vitamin B12. Inactivate. The methionine synthase reductase encoded by MTRR can regenerate functionally active methionine synthase through reductive methylation. MTRR mutation is the main cause of folic acid / methyl vitamin deficiency type E, and it is also one of the main causes of abnormal...

Examples

Embodiment Construction

[0035] The rapid amplification detection kit and detection method of the SNP site of MTRR provided by the present invention will be further described in detail and completely in conjunction with the examples below. The embodiments described below are exemplary only for explaining the present invention and should not be construed as limiting the present invention.

[0036]The experimental methods in the following examples are conventional methods unless otherwise specified. The experimental materials used in the following examples were purchased from the market unless otherwise specified.

[0037] The instruments and reagents adopted in the present invention are as follows:

[0038] PTC-255 PCR amplification instrument, MJ Research, USA;

[0039] Pyrosequencer, Wuhan First Biotechnology Co., Ltd.;

[0040] TaqDNA polymerase, dNTPs (10mmol / L), 10×Buffer and MgCl 2 (25mmol / L) (Dalian TaKaRa company); AmpliTaqGold polymerase (AppliedBiosystems company); KlenowFragment (no exon...