| New buffers to improve the quantitative real-time polymerase chain reaction. | |
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MedLine Citation:
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PMID: 17690445 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Real-time PCR is a potent technique for nucleic acid quantification for research and diagnostic purposes, the wide dynamic range being one of the advantages over other techniques like the microarray. Several additives and enhancers have been studied to expand the PCR dynamic range in order to be more efficient in quantifying low quantities of nucleic acids, increase the yield and improve reaction efficiency. Shown here is that a combination of new buffers with the regularly used Tris buffer makes it possible to expand the real-time PCR dynamic range and to improve the efficiency and correlation coefficient. Mixing HEPES, TEA or MOPS with Tris was more efficient than Tris alone. It was also found that, if the pH value of the Tris buffer was calibrated with phosphoric acid instead of hydrochloric acid, then the dynamic range was significantly improved and low quantities could be detected and quantified more efficiently. Mixing more than one compound with the Tris buffer was also effective for expanding the dynamic range and increasing the efficiency and correlation coefficient in quantitative real-time PCR. |
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Authors:
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Ashraf Ahmad; Jahan Ghasemi |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2007-08-07 |
Journal Detail:
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Title: Bioscience, biotechnology, and biochemistry Volume: 71 ISSN: 0916-8451 ISO Abbreviation: Biosci. Biotechnol. Biochem. Publication Date: 2007 Aug |
Date Detail:
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Created Date: 2007-08-27 Completed Date: 2007-12-20 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9205717 Medline TA: Biosci Biotechnol Biochem Country: Japan |
Other Details:
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Languages: eng Pagination: 1970-8 Citation Subset: IM |
Affiliation:
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Department of Chemical and Biological Engineering-Molecular Biotechnology, Chalmers University of Technology, Göteborg, Sweden. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Buffers* HEPES Hydrogen-Ion Concentration Morpholines Polymerase Chain Reaction / methods*, standards Tromethamine |
| Chemical | |
Reg. No./Substance:
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0/Buffers; 0/Morpholines; 1132-61-2/morpholinopropane sulfonic acid; 7365-45-9/HEPES; 77-86-1/Tromethamine |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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