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Kit for detecting and evaluating genes of children obesity
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A gene detection and kit technology is applied in the field of kits for the detection and evaluation of children's obesity physique to achieve the effect of facilitating popularization and application.
Inactive Publication Date: 2010-03-31
上海裕隆生物科技有限公司
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It should be reminded that the accuracy of this analyzer is related to the measurement time, whether the bladder is emptied, or even whether there is sweat on the soles of the feet, and the error is relatively large (the value of fat children may be low, and the value of thin children may be high) , so it can only be used as a simple measurement, a more accurate judgment, and it must be diagnosed in conjunction with the pediatrics or endocrinology department of the hospital
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Embodiment 1
[0059] Kit preparation. Including (1) probe preparation; (2) DNA extraction reagent; (3) premixed reaction solution; (4) standard positive control; (5) standard negative control.
[0060] (1) Preparation of the probe;
[0061] Synthesized by a common nucleotide primer synthesizer. The sequence is as follows:
[0062] The sequence of the amplification primers for detecting the PPARG gene is as follows:
[0063] Upstream primer F1(A)C 5-TCT GGG AGA TTC TCC TAT TGA CC-3 23bp,
[0064] Upstream primer F1(A)G 5-TCT GGG AGA TTC TCC TAT TGA CG-3 23bp,
[0065] Downstream primer R1 3-TCA AGG AAG GTC TAT GCC GAT AAC-5 24bp;
[0066] The sequence of the amplification primers for detecting the PPARA gene is as follows:
[0067] Upstream primer F2C 5-CAG TAT TGT CGA TTT CAC AAG TTC C-3 25bp,
[0068] Upstream primer F2G 5-CAG TAT TGT CGA TTT CAC AAG TTC G-3 25bp,
[0113] Sampling method: Use a cotton swab to wipe the inside of the cheek 10 times.
[0114] Note: In order to ensure that the sample is not contaminated by food or drink, do not eat or drink within 30 minutes before sampling.
[0115] a) Processing materials: transfer the cotton swab wiped inside the cheeks into a 2ml centrifuge tube, cut the cotton swab part from its stem with scissors, and add 400 μl buffer GA.
[0116] b) 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.
[0117] c) Add 400 μl buffer GB, mix thoroughly by inversion, and place at 70°C for 10 minutes. At this time, the solution should become clear, and briefly centrifuge to remove the liquid droplets on the inner wall of the tube cap.
[0118] d) Add 200 μl of absolute ethanol, mix thoroughly by inversion, and briefly ce...
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Abstract
A kit used for detecting and evaluating genes of children obesity can overcome the limitations of methods for detecting children obesity, such as a BMI measurement method and a fat measurement method,in principles and operation. The kit comprises a DNA extraction reagent, premixed PCR reaction liquid, a standard positive control substance and a standard negative control substance. When the kit isused, the DNA samples of the children to be detected are taken as the detecting objects, after the sample DNA is extracted, the kit is used for reaction on a fluorescence quantitative PCR instrumentand diagnosis is carried out in combination with the reaction results.The method for testing the children obesity provided by the invention is simple and objective, dispenses with fussy multidirectional tests and is free of influence of temporary physical conditions of the children.
Description
technical field [0001] The invention relates to a product for detecting disease susceptibility genes, in particular to a kit for detecting and evaluating children's obesity constitution genes. Background technique [0002] Obesity refers to the excessive storage of human body fat, manifested as an increase in fat cells and (or) cell volume, that is, a state in which adipose tissue blocks throughout the body increase and lose their normal ratio with other tissues. Obesity is a chronic metabolic disease caused by weight gain and excessive fat accumulation caused by genetic and environmental factors. It is an important cause and common cause of hypertension, coronary heart disease, type 2 diabetes, cholecystitis and certain cancers. Pathological basis. There are many causes of obesity, which can be divided into primary (or simple) and secondary obesity according to the cause. There are other clear causes of secondary obesity such as hypothalamic-pituitary infection, tumor, in...
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