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快速PCR方法在金银花真伪鉴别中的应用

Keywords: 快速PCR,金银花,分子鉴定,荧光检测

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Abstract:

建立简单快速的金银花真伪分子鉴别方法。该研究依据金银花trnL-trnF625位G/TSNP位点设计快速位点特异性PCR引物,优化位点特异性PCR条件,对金银花及其9种混淆品进行扩增及检测。当在87℃预变性1min;87℃变性5s,68℃延伸5s,30个循环时,通过加入SYBRGreenI染料染色,金银花样品显绿色荧光,混淆品无荧光。整个PCR反应可在30min内完成。该研究结果说明快速位点特异性PCR能简单快速鉴别金银花及其混淆品。

References

[1]  Yap E P, Mcgee J O. Short PCR product yields improved by lower denaturation temperatures[J]. Nucleic Acids Res, 1991, 19(7):1713.
[2]  Wittwer C T, Fillmore G C, Garling D J. Minimizing the time required for DNA amplification by efficient heat transfer to small samples[J]. Anal Biochem, 1990, 186(2):328.
[3]  Lee H Y, Park M J, Kim N Y, et al. Rapid direct PCR for ABO blood typing[J]. J Forensic Sci, 2011, 56(s1):S179.
[4]  Bergamo E, Chiapolino G, Lignitto L, et al. Evaluation of fast PCR reagents for rapid and sensitive detection of human herpesvirus 8[J]. J Virol Methods, 2012, 181(1):125.
[5]  Zhang L, Dang F, Kaji N, et al. Fast extraction, amplification and analysis of genes from human blood[J]. J Chromatogr A, 2006, 1106(1/2):175.
[6]  袁媛,蒋超,黄璐琦.中药材分子鉴别现场运用的策略与实践[J].中国中药杂志,2013,38(16):2553.
[7]  Wittwer C T, Marshall B C, Reed G H, et al. Rapid cycle allele-specific amplification: studies with the cystic fibrosis delta F508 locus[J]. Clin Chem, 1993, 39(5): 804.
[8]  张文超. 聚合酶链反应(PCR)技术与基因扩增分析仪器(PCR仪)[J]. 生命科学仪器, 2005(3):13.
[9]  杨文超, 张晓东. 快速PCR研究进展[J]. 中国生物工程杂志, 2007(4): 99.
[10]  Hashimoto M, Chen P C, Mitchell M W, et al. Rapid PCR in a continuous flow device[J]. Lab Chip, 2004, 4(6):638.
[11]  Neuzil P, Novak L, Pipper J, et al. Rapid detection of viral RNA by a pocket-size real-time PCR system[J]. Lab Chip, 2010, 10(19):2632.
[12]  Agrawal N, Hassan Y A, Ugaz V M. A pocket-sized convective PCR thermocycler[J]. Angew Chem Int Ed Engl, 2007, 46(23):4316.
[13]  Wittwer C T, Garling D J. Rapid cycle DNA amplification: time and temperature optimization[J]. Biotechniques, 1991, 10(1):76.
[14]  Aboud M, Oh H H, Mccord B. Rapid direct PCR for forensic genotyping in under 25 min[J]. Electrophoresis, 2013, 34(11):1539.
[15]  Yip P Y, Chau C F, Mak C Y, et al. DNA methods for identification of Chinese medicinal materials[J]. Chin Med, 2007, 2(9):9.
[16]  Heubl G U N. New aspects of DNA-based authentication of Chinese medicinal plants by molecular biological techniques[J]. Planta Med, 2010, 76(17):1963.
[17]  蒋超, 张雅华, 陈敏, 等. 基于双向位点特异性PCR的金银花真伪鉴别方法研究[J]. 中国中药杂志, 2012(24):3752.
[18]  蒋超, 黄璐琦, 袁媛, 等. 使用碱裂解法快速提取药材DNA方法的研究[J]. 药物分析杂志, 2013(7):1081.
[19]  钟罗宝, 陈谷. 一种适用于大规模转基因作物PCR检测的简易DNA提取方法[J]. 现代食品科技, 2008(8):794.
[20]  高红, 张志波, 王维林, 等. 双向等位基因特异性PCR快速区分纯合子和杂合子SNP分型的新方法[J]. 医学分子生物学杂志, 2008(6):524.
[21]  French D J, Archard C L, Andersen M T, et al. Ultra-rapid DNA analysis using HyBeaconTM probes and direct PCR amplification from saliva[J]. Mol Cell Probe, 2002, 16(5):319.
[22]  Wernike K, Beer M, Hoffmann B. Rapid detection of foot-and-mouth disease virus, influenza A virus and classical swine fever virus by high-speed real-time RT-PCR[J]. J Virol Methods, 2013, 193(1):50.
[23]  Wheeler E K, Hara C A, Frank J, et al. Under-three minute PCR: probing the limits of fast amplification[J]. Analyst, 2011, 136(18):3707.

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