Bovine thrombin enhances the efficiency and specificity of polymerase chain reaction

The polymerase chain reaction (PCR) has become one of the central techniques in molecular biology since its invention. However, PCR can be fraught with difficulties in various situations, and it is desirable to find novel PCR enhancers suitable for universal applications. Here we show that bovine th...

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Détails bibliographiques
Publié dans:BioTechniques. - 1991. - 57(2014), 6 vom: 11. Dez., Seite 289-94
Auteur principal: Zhang, Ying (Auteur)
Autres auteurs: Li, Xiaoning, Zou, Ruxing, Xue, Ying, Lou, Xinhui, He, Miao
Format: Article en ligne
Langue:English
Publié: 2014
Accès à la collection:BioTechniques
Sujets:Journal Article Research Support, Non-U.S. Gov't PCR bovine serum albumin bovine thrombin enhancer DNA Primers Serum Albumin, Bovine 27432CM55Q Gold plus... 7440-57-5 DNA 9007-49-2 Thrombin EC 3.4.21.5
Description
Résumé:The polymerase chain reaction (PCR) has become one of the central techniques in molecular biology since its invention. However, PCR can be fraught with difficulties in various situations, and it is desirable to find novel PCR enhancers suitable for universal applications. Here we show that bovine thrombin (BT), a well-known coagulation protein, is exceptionally effective at preventing the formation of primer dimers and enhancing the formation of the desired PCR products. The PCR enhancement effects of BT were demonstrated by testing various types of samples, including low-copy synthetic single-stranded DNAs (ssDNAs), synthetic ssDNA pools, human genomic DNA, and hepatitis B virus genomic DNA. In addition, BT was also able to effectively relieve PCR inhibition by nanomaterial inhibitors such as gold nanoparticles (AuNPs) and graphene oxide (GO). Compared with BSA, one of the most popular PCR enhancers, BT was more effective and required concentrations 18-178 times less than that of BSA to achieve a similar level of PCR enhancement
Description:Date Completed 14.05.2015
Date Revised 16.11.2017
published: Electronic-eCollection
Citation Status MEDLINE
ISSN:1940-9818
DOI:10.2144/000114237