Primer group, probe or probe group, method and reagent box and micro-array for predicting alcohol degradation ability and hangover possibility

A primer set and probe set technology, applied in the field of undisclosed energy, can solve the problems of undisclosed alcohol degradation ability and hangover possibility primer set, undisclosed alcohol degradation ability and hangover possibility, etc.

Inactive Publication Date: 2008-04-02
SAMSUNG ELECTRONICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In addition to the above-mentioned conventional techniques, there is no disclosed primer set that can predict the ability of alcohol degradation and the possibility of hangover
There is also no published probe that can predict the ability of alcohol degradation and the possibility of hangover

Method used

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  • Primer group, probe or probe group, method and reagent box and micro-array for predicting alcohol degradation ability and hangover possibility

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0105] Example 1: Selection of primers for predicting alcohol degradation ability and hangover possibility

[0106] In Example 1, primers for predicting alcohol degradation ability and hangover possibility were selected. To this end, the ALDH2 gene sequence, CYP2E1 gene sequence, and ADH2 gene sequence were obtained from Genbank, and DNASTAR program was used to select target sequences for predicting alcohol degradation ability and hangover possibility: exon XII region of ALDH2 gene, 5 '-control region, and exon III region and IX region of ADH2 gene.

[0107] As a result, 7 sets of oligonucleotide primer sets were designed: the oligonucleotide primer set having the sequence described in SEQ ID NO: 1 and 2, the oligonucleotide primer set having the sequence described in SEQ ID NO: 3 and 4 Nucleotide primer set, there is the oligonucleotide primer set of sequence as described in SEQ ID NO:5 and 6, has the oligonucleotide primer set of sequence as described in SEQ ID NO:7 and 8,...

Embodiment 2

[0108] Example 2: Amplification of genes used for predicting alcohol degradation ability and hangover possibility with the primer set of the present invention because

[0109] The 7 sets of primers designed in Example 1 were used to amplify the genes used to predict alcohol degradation ability and hangover possibility.

[0110] 1) Extract the gDNA of the mouth cells

[0111] Saliva was collected from each test subject, from which gDNA was extracted according to the Qiagen DNA Mini Kit protocol. Then, the gDNA was quantitatively analyzed with Picogreen dsDNA reagent (Molecular Probes). For quantitative analysis, gDNA was subjected to gel electrophoresis and sequenced (ABI3700) to determine the degree of degradation and nucleotide sequence of the gDNA.

[0112] 2) Target sequence amplification

[0113] The gDNA prepared in 1) was amplified by PCR using the Cy3-labeled primers of Example 1. PCR was performed using 50 μl of the reaction solution by adding 5 μl of the gDNA ...

Embodiment 3

[0114] Example 3: Detection of genes amplified using the primer set of the present invention on a chip

[0115] In Example 3, the PCR product obtained in Example 2 was allowed to hybridize with the probe immobilized on the chip, and the degree of probe-target hybridization was determined to predict alcohol degradation ability and hangover possibility.

[0116] 1) Manufacturing chips

[0117] For making chips, use SiO 2 Wafer (thickness 1,000 Ȧ). The wafer was coated with a mixture of 20% 3-aminopropylethoxysilane (Sigma) and 5% FC4430, baked in an oven at 120° C. for 40 minutes, and rinsed with triple distilled water.

[0118] As oligonucleotide probe solutions to be used in chips, nucleotides were prepared by dissolving amine-modified nucleotides (80 μM, Bioneer) in 9 mM PEG 10000 (Sigma) and 100% formamide (Sigma) solution (final concentration: 20 μM). Use a microarray spotter (spotter) (Cartesian, pisys 5500) and an SMP3 needle (pin) (Arrayit) to print the nucleotide ...

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Abstract

A primer set and probe set for predicting alcohol-degrading ability and hangover development is provided to improve rapidness and convenience of prediction by amplifying at least one target sequence selected from ALDH2(acetaldehyde dehydrogenase 2) gene, CYP2E1(cytochrome P450 2E1) gene and ADH2(alcohol dehydrogenase 2) gene. An oligonucleotide probe set capable of hybridizable with at least one target sequence selected from exon XII region of ALDH2 gene, 5'-regulating region of CYP2E1 gene and the exon III region and exon IX region of ADH2 gene is selected from (1) an oligonucleotide probe capable of hybridizable with the exon XII region of ALDH2 gene containing 10 or more consecutive nucleotide fragments containing a 11st base in the nucleotide sequence of SEQ ID NO:15, (2) an oligonucleotide probe capable of hybridizable with the 5-regulating region of CYP2E1 gene containing 10 or more consecutive nucleotide fragments containing a 12nd base in the nucleotide sequence of SEQ ID NO:17, (3) an oligonucleotide probe capable of hybridizable with the exon III region of ADH gene containing 10 or more consecutive nucleotide fragments containing a 12nd base in the nucleotide sequence of SEQ ID NO:19, (4) an oligonucleotide probe capable of hybridizable with the exon IX region of ADH2 gene containing 10 or more consecutive nucleotide fragments containing a 12nd base in the nucleotide sequence of SEQ ID NO:21, and (5) an oligonucleotide probe capable of hybridizable with the exon XII region of ADH2 gene containing 10 or more consecutive nucleotide fragments containing a 13rd base in the nucleotide sequence of SEQ ID NO:23.

Description

technical background [0001] The present invention relates to primer sets, probes or probe sets, methods and kits for predicting alcohol degradation ability and hangover potential. More specifically, the present invention relates to a set of oligonucleotide primers for amplifying at least one target sequence selected from the group consisting of aldehyde dehydrogenase 2 (ALDH2), cytochrome P450 2E1 (CYP2E1), and alcohol dehydrogenase 2 (ADH2) , a probe or a probe group specifically hybridizing with the at least one target sequence, a microarray of the probe or the probe group is immobilized, and the probe or the probe group is used to predict alcohol degradation ability and hangover possibility A method, and a kit for predicting alcohol degradation ability and hangover possibility by using the primer set and the probe or probe set. [0002] Description of related fields [0003] The main component of alcoholic beverages is alcohol, which is degraded into acetaldehyde, a subst...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11C12M1/00C12N15/09G01N33/53G01N37/00
CPCC12Q1/6837C12Q1/6876C12Q2563/107
Inventor 白象铉吴祉令金淑荣李贞男郑钟石
Owner SAMSUNG ELECTRONICS CO LTD
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