Analysis of the VNTR locus D1S80 by the PCR followed by high-resolution PAGE.
Analysis of the VNTR locus D1S80 by the PCR followed by high-resolution PAGE.
复制标题
DOI：
--
复制
发表时间：
1991
影响因子：
9.8
通讯作者：
B. Budowle;R. Chakraborty;A. Giusti;A. Eisenberg;R. Allen
中科院分区：
文献类型：
--
作者：
B. Budowle;R. Chakraborty;A. Giusti;A. Eisenberg;R. Allen
D1 S80位点的等位基因数据通过使用PCR和随后的高分辨率、水平PAGE技术和银染色分析获得。与Southern印迹法进行VNTR基因座的RFLP分析相比，该方法具有以下优点：（1）分离等位基因;（2）测量误差最小;（3）对单带VNTR图谱进行正确的基因分型;（4）非同位素分析;（5）电泳分离的永久记录;（6）缩短分析时间。在99个无关的高加索人的样本中，D1 S80基因座表现出37个表型和16个等位基因的杂合性为80.8%。基因型分布符合Hardy-Weinberg平衡的期望值。此外，观察到的等位基因的数量和杂合性的水平，通过这里描述的协议，是一致的，按照一个单一的，同质的，随机交配人口的突变漂移平衡模型的期望与彼此。因此，D1 S80和类似的VNTR基因座的扩增片段长度多态性（AMP-FLP）的分析可能被证明是有用的模型，通过DNA印迹RFLP分型VNTR基因座分析的群体遗传问题。
Allelic data for the D1S80 locus was obtained by using the PCR and subsequent analysis with a high-resolution, horizontal PAGE technique and silver staining. Compared with RFLP analysis of VNTR loci by Southern blotting, the approach described in this paper offers certain advantages: (1) discrete allele resolution, (2) minimal measurement error, (3) correct genotyping of single-band VNTR patterns, (4) a nonisotopic assay, (5) a permanent record of the electrophoretic separation, and (6) reduced assay time. In a sample of 99 unrelated Caucasians, the D1S80 locus demonstrated a heterozygosity of 80.8% with 37 phenotypes and 16 alleles. The distribution of genotypes is in agreement with expected values according to the Hardy-Weinberg equilibrium. Furthermore, the observed number of alleles and the level of heterozygosity, obtained through the protocol described here, were congruent with each other in accordance with the expectation of a mutation-drift equilibrium model for a single, homogeneous, random-mating population. Therefore, the analysis of D1S80 and similar VNTR loci by amplified fragment length polymorphism (AMP-FLP) may prove useful as models for population genetic issues for VNTR loci analyzed by RFLP typing via Southern blotting.