發(fā)布時間：2019-06-10 15:44

【摘要】：目的 探索高效液相色譜對STR進行分型的方法,掌握高效液相色譜對STR分型的一般規(guī)律。方法 選擇一個STR基因座D10S2325,找到其所有的14個等位基因,分別擴增后混合制備分型標準物。優(yōu)化色譜條件(溫度,分析時間),使分型標準物中所有的等位基因得到分離;考察高效液相色譜分析STR的穩(wěn)定性,根據(jù)對比STR在高效液相色譜中保留時間與分型標準物的保留時間進行STR分型,將結(jié)果與聚丙烯酰胺凝膠電泳分型結(jié)果對比,驗證該法對STR分型的準確性;測定該法檢測單個樣本的靈敏度和混合樣本的能力。結(jié)果 找到了優(yōu)化的色譜條件(溫度為50℃,分析時間為8分鐘左右)。我們觀察到連續(xù)同一批樣本分析結(jié)果穩(wěn)定,但是批間差異尤其是在重新配制試劑后的偏差很大,需要與分型標準物同批進行比對。樣本濃度的變化會引起保留時間有規(guī)律的變化,通過建立線性回歸方程對不同的樣本濃度導(dǎo)致的保留時間偏移進行了有效的校正。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，

本文編號：2496558