权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

DNA Fingerprinting by Means of a Nonradioactive Probe for Individual Identification

利用非放射性探针进行 DNA 指纹识别以进行个体识别

基本信息

批准号：
63570270
负责人：
YAMANOUCHI Haruo
金额：
$ 1.34万
依托单位：
Niigata University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1988
资助国家：
日本
起止时间：
1988 至 1990
项目状态：
已结题

项目摘要

A nonradioactive DNA fingerprinting using ECL (Enhanced Chemi-Luminesent) technique was performed. Three to five mug of genomic DNA digested with Hae III was electrophoresed in 1.3% agarose and transferred onto nylon membrane. Following alkali fixation, the blot was hybridized with peroxidase-labeledminisatellite " myo " probe and washed. Blue light araising from ECL reaction catalyzed by the enzyme label was detected on a light sensitive film. A commercially available kit was employed for DNA detection and labeling. Another minisatellite probe could be reprobed without removing myo. This technique had comparable sensitivity to radioactive method in DNA detection, and was far easier to reprobe than other non-isotopic methods using sulfonated, hapten-labeled and biotin-labeled DNA probe.Actually, this method could be applied to paternity testing, zygosity diagnosis and confirmation of bone marrow engraftment. Thus, since the handling of cold probe is safe and not contaminable, this technique is considered feasible for routine work involving individual identification.

使用ECL（增强化学发光）技术进行非放射性DNA指纹分析。将3 - 5 μ g用Hae III消化的基因组DNA在1.3%琼脂糖中消化并转移到尼龙膜上。碱固定后，将印迹与过氧化物酶标记的小卫星“myo”探针杂交并洗涤。在光敏膜上检测到由酶标记催化的ECL反应产生的蓝光。采用市售试剂盒进行DNA检测和标记。另一个小卫星探针可以在不切除肌萎缩侧索的情况下重新探测。该技术的灵敏度与放射性DNA检测法相当，且比磺化法、半抗原标记法和生物素标记法等非同位素DNA检测法更易于重复检测，可用于亲子鉴定、合子诊断和骨髓植入确认。因此，由于冷探针的处理是安全的且不会受到污染，因此认为该技术对于涉及个体识别的常规工作是可行的。