CN105586396A - Detection kit and method for assessing human body temperature change in fitness exercise - Google Patents
Detection kit and method for assessing human body temperature change in fitness exercise Download PDFInfo
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- CN105586396A CN105586396A CN201410651964.2A CN201410651964A CN105586396A CN 105586396 A CN105586396 A CN 105586396A CN 201410651964 A CN201410651964 A CN 201410651964A CN 105586396 A CN105586396 A CN 105586396A
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Abstract
The invention provides a gene detection kit for assessing body temperature change of a human body in fitness exercise. The kit comprises a kit body and reagents which are kept in the kit body in a separated mode, wherein the reagents include the follows: (1) a PCR (polymerase chain reaction) primer group in accordance with SNP (single nucleotide polymorphism) sites of two genes, (2) a PCR amplification reagent and (3) an agarose gel electrophoresis analysis reagent. According to the detection kit and method disclosed by the invention, a plurality of PCR amplification reactions can be simultaneously conducted on a same PCR instrument through the design of PCR primers, the related genes of a tested object are detected and the high efficiency and the specificity of the detection are achieved, and upon analysis, genetic factors of the body temperature change are obtained, so as to offer an appropriate diet management scheme to the tested object; and the exercise intensity and comfort of the tested object are mastered and corresponding strategies are adopted so as to avoid invalid, and even harmful, methods. Meanwhile, scientific basis is provided for professional athletes in material selecting and training.
Description
Technical field
The present invention relates to genetic test, be specifically related to human health while moving body temperature change gene detecting kit.
Background technology
Body temperature is the necessary condition that human body carries out metabolism and normal activities. Body temperature is subject to again various factors, as muscle activity, and the variation of stress and environment temperature. Human body is in the time of rest state, and the heat production of the internal organs such as liver, intestines and kidney is in 50% left and right of body total amount of heat, and the quantity of heat production of breathing, circulation and brain accounts for 30%, and human skeletal muscle's quantity of heat production only accounts for 20% left and right; But skeletal muscle becomes topmost heat production organ while being in motion state, account for 90% of total amount of heat. Traditional view thinks that human body heat production increases because metaboilic level in motion improves, although regulate and strengthened heat radiation process through body, but still can not ensure body temperature balance and makes hyperthermia. Hyperthermia is larger, and physically-draining is faster, also causes strong discomfort simultaneously, affects locomitivity
When different individual movement, body temperature intensity of variation difference, is mainly because the difference of gene causes. Therefore, no matter carry out body building for common crowd, or select and train for professional sports sportsman, body temperature intensity of variation is all an important measurement index.
Summary of the invention
In view of this, the invention provides a kind of body temperature and change gene detecting kit, this kit detects testee's related gene, analyze the body temperature intensity of variation while obtaining its motion, and pcr amplification reaction synchronously carries out and possess high efficiency, the specificity of detection on same PCR instrument.
A kind of body temperature changes gene detecting kit, comprises the reagent of depositing separately in box body and box body, and the reagent of depositing comprises:
(1) react primer sets for the PCR of 2 gene SNP polymorphic sites:
Primer sets for SNPrs2360969:
5’-GTATTTCAACAAGTCTTGCC-3’
5’-GCATCGGAATCACCA-3’
5’-AGCAATGGGTCTCAAAATG-3’
5’-GAAGCAGGAGGAGTC-3’
Primer sets for SNPrs2253206:
5’-GATAAGTTACAGTTAA-3’
5’-CCCATTCCTCCTACC-3’
5’-GACCACTCAGAAATTCAC-3’
5’-TCTCTCCAGCCTCTCC-3’
(2) pcr amplification reagent;
(3) agarose gel electrophoresis analytical reagent.
The beneficial effect that body temperature provided by the invention changes gene detecting kit is: by the design of PCR primer, multiple pcr amplification reactions are synchronously carried out on same PCR instrument, related gene to testee detects, and high efficiency, the specificity of realization detection, analyze body temperature intensity of variation, thereby the scheme that instructs testee should take in the time of body building, avoids the method invalid or even harmful to it. Simultaneously for selection and the training of professional athlete provide scientific basis.
Brief description of the drawings
Fig. 1 is product agarose gel electrophoresis figure.
Detailed description of the invention
Body temperature changes gene detecting kit, comprises the reagent of depositing separately in box body and box body, and the reagent of depositing comprises:
(1) react primer sets for the PCR of 2 gene SNP polymorphic sites:
Primer sets for SNPrs2360969:
5’-GTATTTCAACAAGTCTTGCC-3’
5’-GCATCGGAATCACCA-3’
5’-AGCAATGGGTCTCAAAATG-3’
5’-GAAGCAGGAGGAGTC-3’
Primer sets for SNPrs2253206:
5’-GATAAGTTACAGTTAA-3’
5’-CCCATTCCTCCTACC-3’
5’-GACCACTCAGAAATTCAC-3’
5’-TCTCTCCAGCCTCTCC-3’
2) pcr amplification reagent;
(3) agarose gel electrophoresis analytical reagent.
Below in conjunction with embodiment, method provided by the invention is further described.
The present embodiment adopts body temperature variation gene detecting kit to detect the SNP polymorphic site of 2 genes to tester simultaneously, and according to Genotyping result, analyze body temperature intensity of variation, thereby the scheme that instructs testee should take is avoided the method invalid or even harmful to it in the time of body building. Simultaneously for selection and the training of professional athlete provide scientific basis.
The body temperature using changes the interior reagent of gene detecting kit by forming below:
(1) react primer sets for the PCR of 2 gene SNP polymorphic sites:
Primer sets for SNPrs2360969:
5’-GTATTTCAACAAGTCTTGCC-3’
5’-GCATCGGAATCACCA-3’
5’-AGCAATGGGTCTCAAAATG-3’
5’-GAAGCAGGAGGAGTC-3’
Primer sets for SNPrs2253206:
5’-GATAAGTTACAGTTAA-3’
5’-CCCATTCCTCCTACC-3’
5’-GACCACTCAGAAATTCAC-3’
5’-TCTCTCCAGCCTCTCC-3’
(2) pcr amplification reagent: 10 × PCR buffer solution, this PCR buffer solution is 100mMTris-HClpH8.3,500mMKCl, 15mMMgCl2; DNTPs mixture, this dNTPs mixture is triphosphoric acid guanine deoxyribonucleoside acid dGTP, triphosphoric acid adenyl-deoxyribonucleotide dATP, triphosphoric acid thymidylic acid dTTP, tetra-kinds of nucleotides of triphosphoric acid deoxycytidylic acid dCTP, each concentration of component is 2.5mM; 5U/ μ l thermal starting Taq enzyme.
(3) agarose gel electrophoresis analytical reagent: agarose, TAE buffer solution, DNA molecular amount standard, Goldview dyeing liquor and DNA sample-loading buffer, this buffer solution uses to 1X taking bromophenol blue as indicator dilution.
Using above-mentioned body temperature to change gene detecting kit detects as follows:
(1) extract sample genomic dna.
Gather this tester's saliva, in 1-2ml saliva sample, add 500ul to extract cushioning liquid, this DNA extracts the EDTA of Tris-HClpH7.4,0.5mM and the NaCl of 50mM that cushioning liquid solvent is 50mM, after piping and druming mixes repeatedly, centrifugal 5 minutes of 8000 × g, supernatant discarded, this step repeats once; In the precipitation obtaining, add 500ul lysate, this lysate solvent is 50mMTris-HClpH7.4,50mMTris-HClpH7.4,150mMNaCl, 1mMEDTA, 1%Tritonx-100,1%Sodiumdeoxycholate, 0.1%SDS, Proteinase K 20mg/mL, after the precipitation that thoroughly suspends also fully mixes, room temperature is placed 30 minutes, puts upside down back and forth during this time centrifuge tube for several times; In the mixed liquor obtaining, adding concentration is the aqueous solution 10 μ L of 10mg/mLRNA enzyme, leaves standstill 10min at 37 DEG C; In the supernatant obtaining, add equal-volume benzene phenol-chloroform mixed solution, phenol and chloroform volume ratio are 1: 1, fully mix, and 4 DEG C of mixed liquors, centrifugal 5 minutes of 12000 × g, supernatant moves in clean centrifuge tube; Add equal-volume benzene atmosphere-chloroform-isoamyl alcohol mixed solution, phenol, chloroform and isoamyl alcohol volume ratio are 25: 24: 1, after fully mixing, and 4 DEG C, centrifugal 5 minutes of 12000 × g, supernatant moves in clean centrifuge tube; Add equal-volume chloroform-isoamyl alcohol mixed solution, after fully mixing, 4 DEG C, centrifugal 5 minutes of 12000 × g, supernatant moves in clean centrifuge tube; Add the ice bath aqueous isopropanol of 0.6 times of volume and the 3mol/L SAS of 0.1 times ,-20 DEG C left standstill after 60 minutes, and 4 DEG C, centrifugal 10 minutes of 12000 × g, abandons supernatant; In the sediment obtaining, add 0.5mL70% ethanol washing and precipitating thing, 4 DEG C, centrifugal 5 minutes of 12000 × g, abandons supernatant, and this step repeats once; The sediment natural air drying obtaining, adds the aseptic ultra-pure water back dissolving of 20 μ L, electrophoresis detection ,-20 DEG C of preservations.
(2) pcr amplification
The present embodiment detects rs2360969 and rs2253206 simultaneously. The detection of each SNP needs a pair of upstream and downstream primer, two polymorphic primers totally 4 PCR primers, needs altogether 8 primers. The detection of each SNP need to be done two pipe pcr amplifications, for preventing the reaction of false positive and so on, whether successfully weighs PCR, add a pipe negative control group, be altogether 5 pipe PCR, described negative control group genomic templates and reaction system are the same, but there is no primer.
According to different detection site, prepare respectively PCR reaction system in following ratio with corresponding primer: 2.5 μ l10 × PCR buffer solutions, 2 μ ldNTPs, 0.5 μ l primer sets, thermal starting Taq enzyme 0.15 μ l, genomic templates 80mg, adding ultra-pure water to cumulative volume is 25 μ l.
The Eppendorf test tube that reaction solution is housed is put into ABI9700PCR instrument, reaction condition is set as follows: 95 DEG C of 3min of denaturation; 94 DEG C of 30s of thermal cycle, 60 DEG C of 30s, 72 DEG C of 30s, 35 circulations; Extend 72 DEG C of 3min. After reaction finishes, take out test tube.
Because 5 pipe pcr amplification reactions synchronously carry out on same PCR instrument, in order to realize high efficiency, the specificity of detection, require the Tm value of 8 PCR primers close, need first to carry out a large amount of DNA sequence dna software analysis for this reason and design PCR primer, and carry out the optimization of Tm value and determine the reasonability designing by a large amount of experiments, the detection of the each gene SNP polymorphism of this kit is not only relatively independent but also connect each other, indivisible.
(3) agarose gel electrophoresis detects
Product after amplification is carried out to agarose electrophoresis detection, and process is as follows: 3g agarose, add 100mL deionized water, and microwave-oven-heating 1min, adds 5 μ LGoldview dyeing liquors while being cooled to 60 DEG C, make 3% Ago-Gel. PCR product 10 μ L mix loading with 6 × tbe buffer liquid, 0.8 μ L, voltage 100V electrophoresis 15min. Reading result taking pictures under uviol lamp, gained agarose electrophoresis detects figure as shown in Figure 1.
It is as follows that detection obtains 2 Genotyping analysis results:
| SNP | Somatotype result |
| rs2360969 | CT |
| rs2253206 | GG |
According to above somatotype result, this tester body temperature in the time of motion changes less. Mean that body temperature fluctuating range is little in the time of the motion of large and long period, discomfort is not strong. This tester is applicable to endurance exercise project, has the joyful sense of stronger motion. If professional athlete can strengthen intensity in the time of training, can not cause very strong pain sensation.
The foregoing is only preferred embodiment of the present invention, in order to limit the present invention, within the spirit and principles in the present invention not all, any amendment of doing, be equal to replacement, improvement etc., within all should being included in protection scope of the present invention.
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Claims (8)
1. body temperature changes gene detecting kit, comprises the reagent of depositing separately in box body and box body, it is characterized in that: the reagent of depositing comprises:
(1) react primer sets for the PCR of 2 gene SNP polymorphic sites:
Primer sets for SNPrs2360969:
5’-GTATTTCAACAAGTCTTGCC-3’
5’-GCATCGGAATCACCA-3’
5’-AGCAATGGGTCTCAAAATG-3’
5’-GAAGCAGGAGGAGTC-3’
Primer sets for SNPrs2253206:
5’-GATAAGTTACAGTTAA-3’
5’-CCCATTCCTCCTACC-3’
5’-GACCACTCAGAAATTCAC-3’
5’-TCTCTCCAGCCTCTCC-3’
(2) pcr amplification reagent;
(3) agarose gel electrophoresis analytical reagent.
2. body temperature according to claim 1 changes gene detecting kit, it is characterized in that: described primer consumption is 0.5-1.0 μ l.
3. body temperature according to claim 1 changes gene detecting kit, it is characterized in that: described pcr amplification reagent comprises: 10 × PCR buffer solution, dNTPs, thermal starting Taq enzyme, genomic templates.
4. body temperature according to claim 3 changes gene detecting kit, it is characterized in that: described PCR buffer solution comprises: 100mMTris-HClpH8.3,500mMKCl, 15mMMgCl2。
5. body temperature according to claim 3 changes gene detecting kit, it is characterized in that: described pcr amplification reagent dosage is: 10 × PCR buffer solution, 2.5 μ l, dNTPs2 μ l, thermal starting Taq enzyme 0.1-0.2 μ l, genomic templates 50-100ng.
6. body temperature according to claim 1 changes gene detecting kit, it is characterized in that: described agarose gel electrophoresis analytical reagent comprises: agarose, TAE buffer solution, DNA molecular amount standard, DNA non-toxic dye and DNA sample-loading buffer.
7. body temperature according to claim 6 changes gene detecting kit, it is characterized in that: the preferred BiotiumGelred non-toxic dye of described DNA non-toxic dye.
8. body temperature according to claim 1 changes gene detecting kit, it is characterized in that: for testee provides the detection in when motion body temperature amplitude of variation, amplitude of variation little applicable larger intensity and the motion of long period, the joyful sense of moving is stronger, otherwise, be not suitable for intensity large, the motion that the time is long, more easily agonizes when motion.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112941194A (en) * | 2019-12-11 | 2021-06-11 | 宁波海尔施基因科技有限公司 | Kit and method for detecting genotype of human body-building potential |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1041330A (en) * | 1988-09-20 | 1990-04-18 | 株式会社日立制作所 | Apparatus for controlling elevator |
| US20030138778A1 (en) * | 2001-11-30 | 2003-07-24 | Garner Harold R. | Prediction of disease-causing alleles from sequence context |
| CN102373280A (en) * | 2011-11-03 | 2012-03-14 | 中国人民解放军军事医学科学院微生物流行病研究所 | Anopheles sinensis drug resistance AS-PCR (Polymerase Chain Reaction) detection kit and special primers thereof |
| CN102618626A (en) * | 2011-01-30 | 2012-08-01 | 杭州优思达生物技术有限公司 | Method and kit for rapidly detecting single nucleotide polymorphisms (SNPs) |
| CN103215361A (en) * | 2013-04-18 | 2013-07-24 | 深圳联合医学科技有限公司 | Allele variant detection method, kit and composition |
| CN103911445A (en) * | 2014-03-20 | 2014-07-09 | 刘小芳 | AS-PCR primer design method, gene polymorphism detection method and kit |
-
2014
- 2014-11-17 CN CN201410651964.2A patent/CN105586396A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1041330A (en) * | 1988-09-20 | 1990-04-18 | 株式会社日立制作所 | Apparatus for controlling elevator |
| US20030138778A1 (en) * | 2001-11-30 | 2003-07-24 | Garner Harold R. | Prediction of disease-causing alleles from sequence context |
| CN102618626A (en) * | 2011-01-30 | 2012-08-01 | 杭州优思达生物技术有限公司 | Method and kit for rapidly detecting single nucleotide polymorphisms (SNPs) |
| CN102373280A (en) * | 2011-11-03 | 2012-03-14 | 中国人民解放军军事医学科学院微生物流行病研究所 | Anopheles sinensis drug resistance AS-PCR (Polymerase Chain Reaction) detection kit and special primers thereof |
| CN103215361A (en) * | 2013-04-18 | 2013-07-24 | 深圳联合医学科技有限公司 | Allele variant detection method, kit and composition |
| CN103911445A (en) * | 2014-03-20 | 2014-07-09 | 刘小芳 | AS-PCR primer design method, gene polymorphism detection method and kit |
Non-Patent Citations (1)
| Title |
|---|
| HOLLIS C.KAROLY等: "Genetic Influences on Physiological and Subjective Responses to an Aerobic Exercise Session among Sedentary Adults", 《JOURNAL OF CANCER EPIDEMIOLOGY》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112941194A (en) * | 2019-12-11 | 2021-06-11 | 宁波海尔施基因科技有限公司 | Kit and method for detecting genotype of human body-building potential |
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Application publication date: 20160518 |
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