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  本文選題：法醫(yī)DNA檢驗(yàn) + 國產(chǎn)磁珠法　； 參考：《東北師范大學(xué)》2009年碩士論文

【摘要】： 磁珠法實(shí)用范圍廣,可以用于血斑、唾液斑、精斑、汗斑等大多數(shù)生物樣本的提取,方法操作簡單,不需要特殊的分離設(shè)備,同時也避免了大樣品體系需要反復(fù)離心而造成的DNA損失。本實(shí)驗(yàn)利用一款國產(chǎn)磁珠,在DNA回收純化方面進(jìn)行了一些探索。實(shí)驗(yàn)證明,國產(chǎn)磁珠用于DNA回收純化具有高回收靈敏度、純化效果好的特性,適合在案件檢驗(yàn)中應(yīng)用。 首先,用國產(chǎn)磁珠法、硅珠法和IQ試劑盒分別對不同量的9947標(biāo)準(zhǔn)DNA進(jìn)行提取,使用PowerPlex16試劑盒、ABI9700型擴(kuò)增儀進(jìn)行PCR復(fù)合擴(kuò)增,擴(kuò)增產(chǎn)物應(yīng)用ABI-3100型DNA分析儀檢測。實(shí)驗(yàn)表明,國產(chǎn)磁珠法在熒光定量PCR方面要優(yōu)于硅珠法和IQ試劑盒。然后,用國產(chǎn)磁珠法、硅珠法和IQ試劑盒分別對稀釋30倍、60倍的血液樣本進(jìn)行提取,使用PowerPlex16試劑盒、ABI9700型擴(kuò)增儀進(jìn)行PCR復(fù)合擴(kuò)增,擴(kuò)增產(chǎn)物應(yīng)用ABI-3100型DNA分析儀檢測。結(jié)果表明,國產(chǎn)磁珠法在痕量DNA提取方面要優(yōu)于硅珠法和IQ試劑盒。最后,用國產(chǎn)磁珠法分別對三種污染檢材:帶血跡玻璃、衣領(lǐng)脫落細(xì)胞、牙刷進(jìn)行提取,使用PowerPlex16試劑盒、ABI9700型擴(kuò)增儀進(jìn)行PCR復(fù)合擴(kuò)增,擴(kuò)增產(chǎn)物應(yīng)用ABI-3100型DNA分析儀檢測,16個STR基因座均獲得了理想的DNA分型。結(jié)論,國產(chǎn)磁珠法能夠從含有染料、血紅素、金屬離子等污染成分的樣本中回收純化DNA,并適合復(fù)合STR擴(kuò)增。
[Abstract]:The magnetic bead method has a wide range of applications. It can be used to extract most biological samples such as blood spot, saliva spot, sperm spot, sweat spot and so on. The method is simple to operate and does not require special separation equipment.At the same time, the DNA loss caused by repeated centrifugation of large sample system is avoided.In this experiment, a domestic magnetic beads were used to recover and purify DNA.It is proved by experiments that the domestic magnetic beads have high recovery sensitivity and good purification effect for DNA recovery and purification, and are suitable for application in case testing.Firstly, the 9947 standard DNA was extracted by magnetic bead method, silicon bead method and IQ kit respectively. The PCR was amplified by PowerPlex16 kit ABI9700 amplification instrument, and the amplification product was detected by ABI-3100 DNA analyzer.The results show that the domestic magnetic bead method is superior to the silicon bead method and IQ kit in fluorescence quantitative PCR.Then, the blood samples with 30 times dilution and 60 times dilution were extracted by magnetic bead method, silicon bead method and IQ kit respectively. The PCR was amplified by PowerPlex16 kit ABI9700, and the products were detected by ABI-3100 DNA analyzer.The results showed that the domestic magnetic bead method was superior to the silicon bead method and IQ kit in the extraction of trace DNA.Finally, three kinds of contaminated samples were extracted by domestic magnetic bead method: stained glass, neckline exfoliated cells, toothbrush, and PCR was amplified by PowerPlex16 kit ABI9700.The amplification products were detected by ABI-3100 type DNA analyzer, and all 16 STR loci obtained ideal DNA typing.Conclusion: the domestic magnetic beads method can recover and purify DNA from samples containing dye, heme and metal ions, and is suitable for compound STR amplification.
【學(xué)位授予單位】：東北師范大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2009
【分類號】：D919

【引證文獻(xiàn)】

相關(guān)期刊論文 前2條

1 曲春冰;張國翔;袁家龍;;KingFisher磁珠純化儀在提取脫落細(xì)胞檢材DNA上的應(yīng)用[J];廣東公安科技;2011年01期

2 申萬祥;姚默;趙兵;崔超;鞏江;倪士峰;;生物考古中的DNA分析技術(shù)概況[J];畜牧與飼料科學(xué);2012年02期

相關(guān)碩士學(xué)位論文 前1條

1 張景閣;金磁微粒純化細(xì)菌基因組DNA方法的研究[D];西北大學(xué);2011年

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本文編號：1742679