本文選題：miniSTR + 短串聯(lián)重復(fù)序列��； 參考：《四川大學(xué)》2007年碩士論文

【摘要】： 目的對THO1、TPOX、CSF1PO三個基因座PCR-miniSTR和普通引物PCR-STR的結(jié)果進(jìn)行一致性檢驗(yàn)；構(gòu)建THO1、TPOX、CSF1PO三個基因座miniSTR熒光標(biāo)記復(fù)合擴(kuò)增體系；對該熒光標(biāo)記復(fù)合擴(kuò)增體系進(jìn)行法醫(yī)學(xué)應(yīng)用性研究。方法參照Bulter等對THO1、TPOX、CSF1PO三個基因座miniSTR的引物設(shè)計，選用本教研室提供的123例無血緣關(guān)系個體的血樣，進(jìn)行THO1、TPOX、CSF1PO三個基因座PCR-miniSTR銀染檢測，檢測結(jié)果與商品化試劑盒的檢測結(jié)果進(jìn)行一致性檢驗(yàn)；采用國產(chǎn)DNA聚合酶構(gòu)建miniSTR熒光標(biāo)記復(fù)合擴(kuò)增體系，用美國ABI-310基因分析儀進(jìn)行檢測；對該熒光標(biāo)記復(fù)合擴(kuò)增體系的靈敏度、準(zhǔn)確性、種屬特異性、不同退火溫度下復(fù)合擴(kuò)增的效果、陳舊血痕DNA的檢測等進(jìn)行研究。結(jié)果123例無血緣關(guān)系個體的THO1、TPOX、CSF1PO三個基因座PCR-miniSTR銀染檢測結(jié)果與商品化試劑盒檢測結(jié)果一致；成功建立了THO1、TPOX、CSF1PO三個基因座miniSTR熒光標(biāo)記復(fù)合擴(kuò)增體系，該體系的的退火溫度范圍較寬，以60℃為最佳，檢測靈敏度為0.25ng模板DNA，，擴(kuò)增準(zhǔn)確性高，具有較高的種屬特異性，可以應(yīng)用于陳舊血痕DNA檢測。結(jié)論THO1、TPOX、CSF1PO三個基因座PCR-miniSTR結(jié)果可以與該三個基因座普通引物PCR-STR的結(jié)果進(jìn)行比對，具有數(shù)據(jù)庫兼容性；該熒光標(biāo)記復(fù)合擴(kuò)增體系可應(yīng)用于ABI-310檢測平臺，擴(kuò)增靈敏度高，條件易于優(yōu)化、分型結(jié)果穩(wěn)定，種屬特異性好，可應(yīng)用于法醫(yī)學(xué)實(shí)際檢案。
[Abstract]:Objective to verify the consistency between the PCR-miniSTR of TPOXG CSF1PO and the PCR-STR of common primers, to construct a fluorescent multiplex amplification system for miniSTR of TPOXG / CSF1PO, and to study the forensic application of the system. Methods according to the primer design of miniSTR of three loci of THO1TPOXG CSF1PO, the blood samples of 123 unrelated individuals from our teaching and research department were used to detect the PCR-miniSTR silver staining of the three loci of THO1, TPOX, CSF1PO. The detection results were consistent with those of commercial kits. The miniSTR fluorescent labeling multiplex amplification system was constructed by domestic DNA polymerase and detected by American ABI-310 gene analyzer. The sensitivity of the fluorescent labeling multiplex amplification system was analyzed. Accuracy, species specificity, multiple amplification at different annealing temperatures, and DNA detection of old blood marks were studied. Results the results of PCR-miniSTR silver staining of the three loci of THO1 and TPOXF1PO in 123 unrelated individuals were in agreement with the results of commercial kit, and the fluorescent multiplex amplification system was successfully established for the miniSTR labeling of the three loci of THO1, TPOX, CSF1PO, and the annealing temperature of the system was wide. At 60 鈩

本文編號：1950741