The Application of DNA Polymorphism to Paternity Testing

亲子鉴定后用苯酚-氯仿法或商用DNA提取试剂盒从冷冻保存的淋巴细胞中提取DNA。父权分析已进行了一种简单的方法，非放射性DNA指纹。该方法利用地高辛标记的微型探针MZ1.3，使用抗地高辛-碱性磷酸酶复合物检测。该系统检测了来自淋巴细胞的超过10毫升DNA中的DNA限制性片段。另一方面，本实验通过聚合酶链反应（PCR）扩增HLA DQ - a基因，并随后与序列特异性寡核苷酸探针（Ampli Type kit）杂交，评估HLA DQ - a基因的分型。通过扩增不同数量的纯化DNA来评估检测淋巴细胞DNA提取的灵敏度。从1 ~ 20ng作为亲子鉴定的模板DNA进行HLA DQ分型。

DNA was extracted from cryopreserved lymphocytes after paternity testing by the phenol-chloroform method or commercial DNA extraction kits. Paternity analysis has been performed by a simple method for nonradioactive DNA fingerprinting. The method utilizes a minisatelite probe MZ1.3 labeled with digoxigenin, which are detected using an anti-digoxigenin-alkaline phosphatase complex. This system detected DNA restriction fragments in more than 10mug of DNA from lymphocytes.On the other hand, the experiment has been performed to evaluate typing of the HLA DQ a gene by polymerase chain reaction (PCR) amplification of the gene and subsequent hybridization with sequence-specific oligonucleotide probe (Ampli Type kit). The sensitivity to detect DNA extraction from the lymphocytes was evaluated by amplifying various amounts of purified DNA.HLA DQ a typing were detected from 1-20ng as amounts of template DNA from paternity test.