Boonsaeng V
Department of Biochemistry, Faculty of Science Mahidol University, Bangkok, Thailand.
Southeast Asian J Trop Med Public Health. 1995;26 Suppl 1:296-300.
In forensic medicine, DNA fingerprinting for identification is becoming a necessary procedure. A method to radiolabel M13 DNA probe by primer extension using a specific oligonucleotide primer was developed. The method specifically labeled the two 15 bp repeats in M13 DNA which hybridize to target DNA giving rise to DNA fingerprinting patterns. The M13 probe labeled by this method has proven useful for individual identification, paternity testing and monitoring reconstitution in bone marrow transplantation. The genetic locus D1S80 and D17S30 containing a variable number of tandem repeats (VNTR) have also been successfully amplified from human genomic DNA isolated from blood (50 ng from each sample) by the polymerase chain reaction (PCR) using oligonucleotide primers complementary to the flanking sequences as primers for amplification. DNA bands were detected by ethidium bromide staining after electrophoresis on agarose gels. Analysis of this VNTR locus was thus achieved without the need for Southern blot or radioactive material. The small size of the DNA fragments produced in the PCR amplification permited good resolution of individual alleles. The precise specification of the number of tandem repeats present in each allelic fragment was reproducible from one analysis to another.
在法医学中，用于身份鉴定的DNA指纹识别正成为一项必要的程序。开发了一种使用特定寡核苷酸引物通过引物延伸对M13 DNA探针进行放射性标记的方法。该方法特异性地标记了M13 DNA中与靶DNA杂交的两个15 bp重复序列，从而产生DNA指纹图谱。通过这种方法标记的M13探针已被证明可用于个体识别、亲子鉴定和监测骨髓移植中的重建情况。含有可变数量串联重复序列（VNTR）的遗传位点D1S80和D17S30也已通过聚合酶链反应（PCR）从血液中分离的人类基因组DNA（每个样品50 ng）中成功扩增出来，使用与侧翼序列互补的寡核苷酸引物作为扩增引物。在琼脂糖凝胶上电泳后，通过溴化乙锭染色检测DNA条带。因此，无需Southern印迹或放射性物质即可实现对该VNTR位点的分析。PCR扩增产生的DNA片段尺寸小，使得各个等位基因具有良好的分辨率。每个等位基因片段中串联重复序列数量的精确确定在每次分析之间具有可重复性。
