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Amplification primer, kit and detection method for evaluating folic acid metabolism ability

A technology for folic acid metabolism and primer amplification, which is applied in biochemical equipment and methods, microbiological measurement/inspection, DNA/RNA fragments, etc. It can solve problems such as poor detection accuracy, complicated operation, and experimental failure, and achieve accurate results , simple operation and low cost

Pending Publication Date: 2019-01-11
ALLIFE MEDICAL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

PCR technology has become one of the most widely used methods in molecular biology research, and primer design is a crucial part of PCR technology, and the use of inappropriate PCR primers can easily lead to experimental failure
[0007] The detection methods of folic acid metabolism-related genes in the prior art generally have the defects of poor detection accuracy, long cycle, high cost, and complicated operation

Method used

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  • Amplification primer, kit and detection method for evaluating folic acid metabolism ability
  • Amplification primer, kit and detection method for evaluating folic acid metabolism ability
  • Amplification primer, kit and detection method for evaluating folic acid metabolism ability

Examples

Experimental program
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Effect test

Embodiment 1

[0052] An amplification primer for evaluating folic acid metabolism ability, comprising a first primer pair for detecting rs1801133, a second primer pair for detecting rs1801131, a third primer pair for detecting rs1801394 and a fourth primer pair for detecting rs1805087 Right, wherein said first primer pair includes the nucleotide sequence shown in SEQ ID NO.1 and SEQ ID NO.2, and said second primer pair includes the nucleotide sequence shown in SEQ ID NO.3 and SEQ ID NO.4 Nucleotide sequence, the third primer pair includes the nucleotide sequence shown in SEQ ID NO.5 and SEQ ID NO.6, and the fourth primer pair includes the nucleotide sequence shown in SEQ ID NO.7 and SEQ ID NO.8 the nucleotide sequence.

[0053] The sequence list of the above-mentioned amplification primers for evaluating folic acid metabolism ability is shown in Table 1.

[0054] Table 1 sequence list

[0055]

Embodiment 2

[0057] A kit for evaluating folic acid metabolism ability, said kit comprising the above-mentioned amplification primers for evaluating folic acid metabolism ability.

[0058] The kit also includes PCR buffer, dNTPs, Mg 2+ , 2XTaq enzyme and reagents for extracting oral mucosal cell DNA from the sample to be tested.

[0059] The reagent used to extract the DNA of oral mucosal cells of the sample to be tested was purchased from Tiangen Biochemical Technology (Beijing) Co., Ltd., and the model was oral swab genomic DNA extraction kit (DP322)

Embodiment 3

[0061] A method for evaluating folic acid metabolism capacity, comprising the steps of:

[0062] Step 1: Extract DNA from oral mucosal cells of the sample to be tested

[0063] Tiangen Biochemical Technology (Beijing) Co., Ltd., the model is the oral swab genomic DNA extraction kit (DP322), which extracts DNA from the oral mucosal cells of the tester.

[0064] Step 2: use the nucleotide sequence shown in SEQ ID NO.1 and SEQ ID NO.2 to amplify the MTHFR gene C677T respectively, and use the nucleotide sequence shown in SEQ ID NO.3 and SEQ ID NO.4 to amplify MTHFR gene A1298C, use the nucleotide sequence shown in SEQ ID NO.5 and SEQ ID NO.6 to amplify MTRR gene A66G, use the nucleotide sequence shown in SEQ ID NO.7 and SEQ ID NO.8 to amplify MTR For the gene A2756G, the DNA extracted from the oral mucosal cells obtained in step 1 was used as a template, and PCR reactions were performed respectively, and 20uL systems were taken respectively, including:

[0065] 50ng / uL of templa...

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PUM

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Abstract

The invention discloses an amplification primer, a kit and a detection method for evaluating folic acid metabolism ability, belonging to the field of biotechnology. The amplification primer for evaluating folic acid metabolism ability comprises a first primer pair for detecting rs1801133, a second primer pair for detecting rs1801131, a third primer pair for detecting rs1801394 and a fourth primerpair for detecting rs1805087. The invention also discloses a kit for evaluating folic acid metabolism ability and a method for evaluating folic acid metabolism ability. The amplification primer for evaluating folic acid metabolism ability has the advantages of high sensitivity, high specificity, high accuracy and simple operation, and can realize gene detection of four polymorphic sites related tofolic acid metabolism, screen out mutation sites, realize individualized supplementation of folic acid, and reduce the risk of birth defects of newborns.

Description

technical field [0001] The invention relates to an amplification primer, a kit and a detection method for evaluating folic acid metabolism ability, and belongs to the field of biotechnology. Background technique [0002] Folic acid, also known as vitamin B9, is an essential substance in the growth process of the fetus. It is a nutrient in the vitamin B group and is water-soluble, so it cannot be gathered in the body. This nutrient was first discovered in spinach leaves, hence the name "folate". Folic acid plays an important role in the division and growth of cells and the synthesis of nucleic acids, amino acids and proteins. The lack of folic acid in the human body can lead to abnormal red blood cells, an increase in immature cells, anemia and a decrease in white blood cells. Folic acid is also an indispensable nutrient for fetal growth and development. Folic acid deficiency in pregnant women may lead to low birth weight, cleft lip and palate, and heart defects. If folic...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/156
Inventor 顾雨春宋娜
Owner ALLIFE MEDICAL SCI & TECH CO LTD
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