CAJ | 학술논문

为寻求一种从藏酋猴毛干中提取总DNA的有效方法,从一藏酋猴个体背部剪取数量不等的毛干样品(不含毛囊),经蛋白酶K消化后,分别采用高盐沉淀抽提法、酚氯仿抽提法、纳米磁珠抽提法分离获得用于PCR扩增的模板DNA,通过紫外分光光度计测定OD值并计算出质量浓度,同时进行琼脂糖凝胶电泳和PCR扩增检测,分析3种方法提取藏酋猴毛干DNA的质量差异.每处理设置10个重复.结果表明,纳米磁珠法提取藏酋猴毛干中DNA是一种快捷、简便、经济、高效、安全的有效方法;进行毛发取样时以5~10根较为合适.
위심구일충종장추후모간중제취총DNA적유효방법,종일장추후개체배부전취수량불등적모간양품(불함모낭),경단백매K소화후,분별채용고염침정추제법、분록방추제법、납미자주추제법분리획득용우PCR확증적모판DNA,통과자외분광광도계측정OD치병계산출질량농도,동시진행경지당응효전영화PCR확증검측,분석3충방법제취장추후모간DNA적질량차이.매처리설치10개중복.결과표명,납미자주법제취장추후모간중DNA시일충쾌첩、간편、경제、고효、안전적유효방법;진행모발취양시이5~10근교위합괄.
In order to find an effective method to extract DNA from the hair shaft of Macaca thibetana, different numbers of hair shafts without bottom follicle were sampled from dorsal area of a tibetan macaque, which were digested with SDS-proteinase K and extracted by high salt method, phenol-chloroform method and magnetic bead method. As a result, total genomic DNAs to be used as templates for PCR amplification were obtained. Ten repetitions per sample were set in the present study. The extraction results of the three methods were detected through agarose gel electrophoresis and PCR amplification, and optical density value was determined with an ultraviolet spectrophotometer and the content of DNA was calculated. The analysis result indicates that the magnetic bead extraction method is a quick, simple, economical and efficient method to extract DNA from the hair shaft of tibetan macaque. In addition, 5~10 hair shaft samples should be used in the experiment.