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目的探讨对法医STR基因分型检测使用的毛细管电泳筛分介质进行评价的参数,为电泳凝胶验证评价提供参考指标。方法采用ABI 3130xl遗传分析仪,以3130 POP-4TM凝胶为筛分介质,以分子量内标GS500LIZ和Typer500为样本,多批次进行毛细管电泳;对原始数据进行分组统计,得到单碱基对相对迁移时间及其相对标准偏差(RSD)。结果分子量内标GS500LIZ和Typer500在POP-4TM凝胶中电泳,单碱基对相对迁移时间的统计结果表现趋势相似,即随着DNA片段增大,单碱基对相对迁移时间的均值减小,标准差及相对标准偏差增大。组间方差分析结果符合方差齐性,P<0.01,组间多重比较,P<0.000 28(Bonferron校正)。结论单碱基对相对迁移时间的相对标准偏差(RSD)是毛细管电泳重现性的重要参数,可为凝胶研发和验证提供参考性指标。
OBJECTIVE: To evaluate the parameters of capillary electrophoresis screening media used in forensic STR genotyping test, and to provide a reference index for electrophoresis gel validation. Methods The ABI 3130xl genetic analyzer was used to screen 3130 POP-4TM gels. The molecular weight internal standard GS500LIZ and Typer500 were used as samples. The raw data were grouped by capillary electrophoresis, and the single base pair was obtained. Migration time and its relative standard deviation (RSD). Results The molecular weight of internal standard GS500LIZ and Typer500 in POP-4TM gel electrophoresis, single base pairs of the relative migration time of the statistical results showed a trend similar to that with the increase of DNA fragments, single base pairs relative migration time mean decreases, Standard deviation and relative standard deviation increase. Analysis of variance between groups was consistent with homogeneity of variance, P <0.01, multiple comparisons between groups, P <0.00028 (Bonferron correction). Conclusion The relative standard deviation (RSD) of single-base pair relative migration time is an important parameter for the reproducibility of capillary electrophoresis. It can provide a reference index for gel development and validation.