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Molecular marker kit for formulating individual weight reduction scheme

A technology of molecular markers and kits, applied in the field of kits, can solve the problems of high cost, expensive instruments, waste of SNP site typing, etc., and achieve the effect of low cost, high sensitivity, and large-scale detection

Inactive Publication Date: 2012-09-19
CHONGQING KANONG TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0020] At present, commercial genotyping methods mainly include: 1) Although Microarray chips can detect genotypes with high throughput and more accurately, but the cost is high, and the instrument is expensive; Typing, but the machine is expensive and the cost is high. At the same time, it is obviously too wasteful to use the specific SNP locus typing
3) Although the fluorescent PCR system is more economical than the previous two methods, the detection scale is limited, which is not conducive to commercialization

Method used

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  • Molecular marker kit for formulating individual weight reduction scheme
  • Molecular marker kit for formulating individual weight reduction scheme
  • Molecular marker kit for formulating individual weight reduction scheme

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1 : DNA extraction of oral mucosal cells

[0056] The mucosal cells in the subject's oral cavity were collected with an oral mucosal swab, and the genomic DNA of the subject was extracted using the oral swab genomic DNA extraction kit produced by Shanghai Tiangen Biotechnology. Proceed as follows:

[0057] 1. Processing materials:

[0058] Transfer the swab wiped in the cheeks to a 2ml centrifuge tube, cut the cotton swab part from its stem with scissors, and add 400 μl buffer GA.

[0059] 2. Add 20 μl of proteinase K solution, vortex for 10 seconds to mix, and place at 56°C for 60 minutes, during which time, vortex and mix several times every 15 minutes.

[0060] 3. Add 400 μl buffer GB, mix thoroughly by inverting, and place at 70°C for 10 minutes. At this time, the solution should become clear. Centrifuge briefly to remove the droplets on the inner wall of the tube cap.

[0061] 4. Add 200 μl of absolute ethanol, mix thoroughly by inversion, and briefly ...

Embodiment 2

[0069] Example 2 : PCR amplification of fragments containing specific SNP sites

[0070] 1. Preparation of primers: SEQ ID NO: 1-10 in sequence

[0071] The following primers No. 1 to No. 5 were synthesized by Shenzhen Huada Genomics:

[0072] Table 1

[0073]

[0074] Ultrapure water is required. According to the quality of the primers, add water to dilute the upstream and downstream primers to a concentration of 10 μmol·L-1, and store at 4°C.

[0075]The above five pairs of primers are based on the gene sequence published by NCBI, and the sequence numbers of the genes are FTOrs9939609, FABP2 rs1799883, ADRB3 rs4994, PPARG rs1801282, ADRB2rs1042714. Among them, SEQ NO: 1-2, 7-10 were designed using the software oligo 6.0; SEQ NO: 3-4 came from the literature: Peng Xian'e, Zhang Ling, Wang Qingqing, etc. Research on the association between FABP2 Ala54 Thr gene polymorphism and non-alcoholic fatty liver. Health Research. 2009, 4: 401-404; SEQ NO: 5-6 from literature: El...

Embodiment 3

[0087] Example 3 : SNP typing of the target gene

[0088] SNP analysis was carried out on the target fragments SEQ ID NO: 11-15 of the five genes obtained above using different methods, and the rs1799883 site of the FABP2 gene, the rs4994 site of the ADRB3 gene, and the rs4994 site of the ADRB2 gene were respectively analyzed by enzyme digestion. The rs1042714 site was typed; the rs9939609 site of the FTO gene and the rs1801282 site of the PPARG gene were typed by PCR-SSCP method.

[0089] 1. rs1799883 locus typing of FABP2 gene

[0090] The PCR amplification fragment of FABP2 gene is as follows: SEQ ID NO: 12

[0091] 1 ACAGGTGTTA ATATAGTGAA AAGGAAGCTT GCAGCTCATG ACAATTTGAA GCTGACAATT

[0092] 61 ACACAAGAAG GAAATAAATT CACAGTCAAAA GAATCAAGC N CTTTTCGAAA CATTGAAGTT

[0093] 121 GTTTTTGAAC TTGGTGTCAC CTTTAATTAC AATTCTAGCAG ACGGAACTGA ACTCAGGGTA

[0094] 181 AGAATTTTTT TTTTTATGAG CAATGCATTC TTGATTTTTC TACCCAATAT TAAAATGATT

[0095] 241 TCTGCTCTAT TTCATTGGAT GGTTTAATTA AT...

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Abstract

The invention relates to a molecular marker kit for formulating an individual weight reduction scheme. The kit comprises nucleotide shown in SEQ ID NO:1-10 and can comprise restriction endonucleases Hha I, Msp I and Bbv I. The kit is used for analyzing SNPs (single nucleotide polymorphisms) genotypes of a subject and issuing a comprehensive genetic predisposition analysis report, and nutritionists are invited to recommend a diet style (low fat diet, low carbohydrate diet and balanced diet) suitable to weight reduction to the subject, analyze intake calorie of the subject required every day according to the practical situation of the subject and individually arrange the proportion of three nutrient elements (fat, protein and carbohydrate) in three meals a day, so that healthy, scientific and effective weight reduction is achieved.

Description

technical field [0001] The invention relates to a kit, in particular to a molecular marker kit for formulating an individualized weight loss program. Background technique [0002] In February 2007, the World Health Organization released a survey report on the proportion of obesity in various countries. The report shows that among the 6.5 billion people in the world today, about 1.7 billion people are overweight, of which 300 million are obese. According to statistics, there are currently 200 million overweight people in China, of which more than 90 million are obese. Experts predict that the number of obese people in China will exceed 200 million in the next ten years. [0003] Over the past few decades, the worldwide epidemic of obesity has been fueled by lifestyle changes driven by declining levels of physical activity and overabundance of food. However, the fact that not everyone becomes obese despite being in the same environment suggests that genetics also play a rol...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 鲁浪
Owner CHONGQING KANONG TECH
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