| Temperature-assisted cyclic hybridization (TACH): an improved method for supercoiled DNA hybridization. | |
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MedLine Citation:
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PMID: 20238183 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Accurate hybridization is dependent on the ratio between sequence-specific and unspecific binding. Dissociation of unspecifically bound, while maintaining specifically hybridized, nucleic acids are key steps to obtain a well-defined complex. We have developed a new method, temperature-assisted, cyclic hybridization (TACH), which increases cognate binding at the expense of unspecific hybridization. The method was used for optimizing binding of peptide nucleic acid (PNA) to supercoiled plasmids and has several advantages over previous methods: (1) it reduces the required amount of bis-PNA by three- to fourfold; (2) it results in less unspecific binding; (3) it extends cooperative hybridization, from 3 bp to 5 bp between two adjacent binding sites; and (4) it decreases the aggregation of bis-PNA. This method might be extended to other forms of hybridizations including the use of additional nucleic acids analogs, such as locked nucleic acid (LNA) and, also, to other areas where PNAs are used such as fluorescence in situ hybridization (FISH), microarrays, or in vivo plasmid delivery. |
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Authors:
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Iulian I Oprea; Oscar E Simonson; Pedro M D Moreno; Joana R Viola; Karin E Lundin; C I Edvard Smith |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Molecular biotechnology Volume: 45 ISSN: 1559-0305 ISO Abbreviation: Mol. Biotechnol. Publication Date: 2010 Jun |
Date Detail:
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Created Date: 2010-05-05 Completed Date: 2010-08-10 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9423533 Medline TA: Mol Biotechnol Country: United States |
Other Details:
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Languages: eng Pagination: 171-9 Citation Subset: IM |
Affiliation:
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Department of Laboratory Medicine, Clinical Research Center, Karolinska Institutet, Karolinska University Hospital Huddinge, SE-141 86 Huddinge, Sweden. iulian.oprea@ki.se |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Analysis of Variance Animals DNA, Superhelical / chemistry*, metabolism Electrophoresis, Agar Gel Gene Expression Hydrogen-Ion Concentration Mice Nucleic Acid Conformation Nucleic Acid Hybridization / methods* Peptide Nucleic Acids / chemistry, metabolism Plasmids / chemistry*, metabolism Sensitivity and Specificity Temperature |
| Chemical | |
Reg. No./Substance:
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0/DNA, Superhelical; 0/Peptide Nucleic Acids |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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