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OPRM1基因PCR产物和测序产物三种纯化方法的比较 被引量：2

Comparing Analysis of Three Different Purification Methods for PCR and Sequencing Pruducts in OPRM1 Gene Sequencing
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摘要 目的探讨一种快速、简便、效果好的OPRM1基因PCR产物和测序产物纯化方法，建立稳定的DNA测序技术平台。方法采用乙醇／醋酸钠、过柱和SAp-Exonuclease1结合BigDyeXterminatorPurificationKit3种纯化方法，对50份OPRMl基因PCR产物和测序产物进行纯化效果的比较。结果SAP—ExonucleaseI法对PCR反应产物进行纯化的回收率显著高于乙醇／醋酸钠和过柱法，分别为0．86±0．01，0．48±0．08，0．65±0．01（t=32．5，P〈0．001；t=86．1，P〈0．001）；BigDyeXterminatorPurificationKit法对测序产物进行纯化成功率显著高于乙醇／醋酸钠和过柱法，分别为100％，62％和84％（x2=23．46，P〈0．001；x2=8．70，P=0．003）。结论SAP—ExonucleaseI结合BigDyexterminatorPurifica—tionKit是一种快速、简便、效果好的测序纯化方法。 Objective Looking for a quick,simple and effective purification method for PCR and sequencing product to establish stable DNA sequencing technology platform. Methods 50 PCR products and sequencing products of OPRM1 gene were purified by ethanl/sodium acetate,spin-column and SAP-Exonuelease I combined with BigDye Xterminator Purification Kit. Results The recovery rate of PCR products that was purified by SAP-Exonuelease I methods was significantly higher than by ethanl/sodium aeetate and spin-column methods. Their recovery rates were 0. 86 ± 0. 01,0. 48 ± 0. 08,0. 65 ± 0.0l （ t= 32.5, P〈0. 001 ; t=86.1, P〈0. 001）. And the success rates of sequencing products that was purified by BigDye Xterminator Purification Kit was signifieantly higher than by ethanl/sodium acetate and spin-column. Their success rates were 100%, 62 % and 84 % （X2 = 23.46, P〈 0. 001, x2 = 8.70, P= 0. 003）. Conclusion The method of SAP-Exonuelease I eombined with BigDye Xterminator Purification kit is a quick,easy and effective purification method.
作者 李少英 马晓燕 张双全 张慧敏 黎青
机构地区 广州医学院第三附属医院妇产科
出处 《现代检验医学杂志》 CAS 2012年第1期120-122,共3页 Journal of Modern Laboratory Medicine
基金 基金项目：广东省科技计划项目,项目编号：20098030801351.
关键词 脱氧测序法 虾碱性磷酸酶 核酸外切酶I 阿片受体基因 sanger dideoxy DNA sequencing SAP （shrimp alkaline phosphatase） exonuclease I OPRM1
分类号 Q503 [生物学—生物化学]
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参考文献12

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共引文献4

同被引文献21

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现代检验医学杂志
2012年 第1期

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