| A microarray-based method to perform nucleic acid selections. | |
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MedLine Citation:
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PMID: 20857369 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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This method describes a microarray-based platform to perform nucleic acid selections. Chemical ligands to which a nucleic acid binder is desired are immobilized onto an agarose microarray surface; the array is then incubated with an RNA library. Bound RNA library members are harvested directly from the array surface via gel excision at the position on the array where a ligand was immobilized. The RNA is then amplified via RT-PCR, cloned, and sequenced. This method has the following advantages over traditional resin-based Systematic Evolution of Ligands by Exponential Enrichment (SELEX): (1) multiple selections can be completed in parallel on a single microarray surface; (2) kinetic biases in the selections are mitigated since all RNA binders are harvested from an array via gel excision; (3) the amount of chemical ligand needed to perform a selection is minimized; (4) selections do not require expensive resins or equipment; and (5) the matrix used for selections is inexpensive and easy to prepare. Although this protocol was demonstrated for RNA selections, it should be applicable for any nucleic acid selection. |
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Authors:
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Olga Aminova; Matthew D Disney |
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Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Methods in molecular biology (Clifton, N.J.) Volume: 669 ISSN: 1940-6029 ISO Abbreviation: Methods Mol. Biol. Publication Date: 2010 |
Date Detail:
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Created Date: 2010-09-21 Completed Date: 2011-01-07 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9214969 Medline TA: Methods Mol Biol Country: United States |
Other Details:
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Languages: eng Pagination: 209-24 Citation Subset: IM |
Affiliation:
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Department of Chemistry and The Center for Excellence in Bioinformatics and Life Sciences, University at Buffalo, Buffalo, NY, USA. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Amines
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chemistry Aminoglycosides / chemistry Animals Base Sequence DNA / chemistry, genetics, metabolism Ligands Nucleic Acid Hybridization Nucleic Acids / chemistry, genetics, metabolism* Oligonucleotide Array Sequence Analysis / methods* Oxidation-Reduction RNA / chemistry, genetics, metabolism Reverse Transcriptase Polymerase Chain Reaction Sepharose / chemistry Surface Properties Transcription, Genetic |
| Grant Support | |
ID/Acronym/Agency:
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GM079235/GM/NIGMS NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Amines; 0/Aminoglycosides; 0/Ligands; 0/Nucleic Acids; 63231-63-0/RNA; 9007-49-2/DNA; 9012-36-6/Sepharose |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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