高效液相色谱对STR分型的研究
发布时间：2019-06-10 15:44
【摘要】：目的 探索高效液相色谱对STR进行分型的方法,掌握高效液相色谱对STR分型的一般规律。方法 选择一个STR基因座D10S2325,找到其所有的14个等位基因,分别扩增后混合制备分型标准物。优化色谱条件(温度,分析时间),使分型标准物中所有的等位基因得到分离;考察高效液相色谱分析STR的稳定性,根据对比STR在高效液相色谱中保留时间与分型标准物的保留时间进行STR分型,将结果与聚丙烯酰胺凝胶电泳分型结果对比,验证该法对STR分型的准确性;测定该法检测单个样本的灵敏度和混合样本的能力。结果 找到了优化的色谱条件(温度为50℃,分析时间为8分钟左右)。我们观察到连续同一批样本分析结果稳定,但是批间差异尤其是在重新配制试剂后的偏差很大,需要与分型标准物同批进行比对。样本浓度的变化会引起保留时间有规律的变化,通过建立线性回归方程对不同的样本浓度导致的保留时间偏移进行了有效的校正。20
[Abstract]:Objective to explore the method of STR typing by high performance liquid chromatography (HPLC) and to master the general rule of STR typing by HPLC. Methods A STR locus D10S2325 was selected and all 14 alleles were found. After amplification, the typing standard was prepared. The chromatographic conditions (temperature, analysis time) were optimized to isolate all the alleles in the typing standard. The stability of STR by high performance liquid chromatography (HPLC) was investigated. STR typing was carried out according to the retention time of STR in high performance liquid chromatography (HPLC) and the retention time of typing standard. The results were compared with those of polyacrylamide gel electrophoresis (page). The accuracy of this method for STR typing was verified. The sensitivity of the method and the ability of mixed samples were measured. Results the optimized chromatographic conditions were found (the temperature was 50 鈩，
本文编号：2496558
[Abstract]:Objective to explore the method of STR typing by high performance liquid chromatography (HPLC) and to master the general rule of STR typing by HPLC. Methods A STR locus D10S2325 was selected and all 14 alleles were found. After amplification, the typing standard was prepared. The chromatographic conditions (temperature, analysis time) were optimized to isolate all the alleles in the typing standard. The stability of STR by high performance liquid chromatography (HPLC) was investigated. STR typing was carried out according to the retention time of STR in high performance liquid chromatography (HPLC) and the retention time of typing standard. The results were compared with those of polyacrylamide gel electrophoresis (page). The accuracy of this method for STR typing was verified. The sensitivity of the method and the ability of mixed samples were measured. Results the optimized chromatographic conditions were found (the temperature was 50 鈩，
本文编号：2496558
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