The Application of DNA Polymorphism to Paternity Testing

用苯酚-氯仿法或商品DNA提取试剂盒对超低温保存的淋巴细胞进行亲子鉴定后提取DNA。亲子关系分析是通过一种简单的非放射性DNA指纹分析方法进行的。该方法利用地高辛标记的微型卫星探针MZ1.3，用抗地高辛-碱性磷酸酶复合体检测。该系统从10多个淋巴细胞的DNA中检测到DNA限制性片段，另一方面，通过聚合酶链式反应(PCR)扩增基因并随后与序列特异性寡核苷酸探针(Ampli Type Kit)杂交，对HLADQ a基因进行了分型。通过扩增不同数量的纯化DNA来评价检测淋巴细胞DNA提取的敏感性。从亲子鉴定中检测到1-20 ng的模板DNA作为HLADQa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.