A method for detecting obesity gene, genomic DNA was extracted from the peripheral blood, using 190ngDNA as template, PCR amplification was carried out in the total reaction volume 50ul, PCR reaction condition of 96 DEG C predegeneration 10min; 94 40s denaturation renaturation; 40s; extension of 60 C, 71 C, 1min, 30 cycles, the last one. 72 Ring C extension 5min. The PCR products were electrophoretic by 2% agarose gel, and the fluorescent band in 210b was identified as a specific PCR product under the ultraviolet light transmission lamp. The restriction endonuclease BstNI was added to the PCR amplification products, 3h was cut at 61, and the results were observed with 3% agarose agaramine electrophoresis and UV lamp.
【技术实现步骤摘要】
一种肥胖基因的检测方法
本专利技术涉及一种肥胖基因的检测方法,具体地说是一种肥胖基因的检测方法。
技术介绍
目前公知的肥胖症(obesity)又名肥胖病。肥胖症是一些社会性慢性疾病。机体内热量的摄入量高于消耗,造成体内脂肪堆积过多,导致体重超标、体态臃肿,实际测量体重超过标准体重>20%,且脂肪百分比>30%者称为肥胖。单纯性肥胖是各种肥胖最常见的一种,约占肥胖人群的95%左右。单纯性肥胖又分为体质性肥胖和过食性肥胖两种,续发性肥胖是由内分泌混乱或代谢障碍引起的一类疾病肥胖。
技术实现思路
诊断标准:研究对象一般资料:采用整群抽取的方法,随机调查197名儿童情况,其中男童100名,女童97名,年龄3~6岁。方法;体表测量指标:由专业人员进行体重、身高、皮褶厚度(肱二头肌、肩胛下角),计算体质量指数(BMI)=体重(kg)/身高(m),根据WHO标准分类为正常儿童与肥胖儿童。儿童肥胖诊断标准根据国际肥胖工作组推荐的儿童肥胖诊断标准的BMI年龄图,β-AR多态性检测:常规酚/氯仿抽提外周血基因组DNA,以190ngDNA为模板,在总反应体积50ul进行PCR扩增,PCR反应条件96℃预变性10min;94℃变性40s;复性60℃,40s;延伸71℃,1min,循环30次,最后一个循环72℃延伸5min。PCR产物经2%琼脂糖凝脉电泳,紫外线透射灯下检测210b处出现荧光带为特异性PCR产物。在PCR扩增产物中加入限制性内切酶BstNI,61℃酶切3h,酶切产物用3%琼脂糖凝胺电泳,紫外灯下观察结果。统计分析:采用SPSS11.0统计软件 ...
【技术保护点】
一种肥胖基因的检测方法,抽提外周血基因组DNA,以190ngDNA为模板,在总反应体积50 ul进行PCR扩增, PCR反应条件96℃预变性10min;94℃变性40s;复性60℃,40s;延伸71℃,1min,循环30次,最后一个循环72℃延伸5min;PCR产物经2%琼脂糖凝脉电泳,紫外线透射灯下检测210b处出现荧光带为特异性PCR产物;在PCR扩增产物中加入限制性内切酶BstNI,61℃酶切3h,酶切产物用3%琼脂糖凝胺电泳,紫外灯下观察结果。
【技术特征摘要】
1.一种肥胖基因的检测方法,抽提外周血基因组DNA,以190ngDNA为模板,在总反应体积50ul进行PCR扩增,PCR反应条件96℃预变性10min;94℃变性40s;复性60℃,40s;延伸71℃,1min,循环30次,最后一个循环72℃延伸5min;PCR产物经2%琼脂糖凝脉电泳,紫外线透射灯下检测210b处出现荧光带为特异性PCR产物;在PCR扩增产物中...
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