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  Fluorescence-activated cell sorting for aptamer SELEX with cell mixtures

Mayer, G., Ahmed, M. S., Dolf, A., Endl, E., Knolle, P. A., & Famulok, M. (2010). Fluorescence-activated cell sorting for aptamer SELEX with cell mixtures. Nature Protocols, 5(12), 1993-2004. doi:10.1038/nprot.2010.163.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0028-6435-9 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0028-6436-7
Genre: Journal Article

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Mayer-2010-Fluorescence-activat.pdf (Any fulltext), 657KB
 
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Mayer, G., Author
Ahmed, M. S., Author
Dolf, A., Author
Endl, E., Author
Knolle, P. A., Author
Famulok, M.1, Author
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1External Organizations, ou_persistent22              

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Free keywords: Cell Separation/*methods Flow Cytometry/*methods Gene Library SELEX Aptamer Technique/*methods Tumor Cells, Cultured
 Abstract: Aptamers that target a specific cell subpopulation within composite mixtures represent invaluable tools in biomedical research and in the development of cell-specific therapeutics. Here we describe a detailed protocol for a modular and generally applicable scheme to select aptamers that target the subpopulations of cells in which you are interested. A fluorescence-activated cell-sorting device is used to simultaneously differentiate and separate those subpopulations of cells having bound and unbound aptamers. There are fewer false positives when using this approach in comparison with other cell-selection approaches in which unspecific binding of nucleic acids to cells with reduced membrane integrity or their unselective uptake by dead cells occurs more often. The protocol provides a state-of-the-art approach for identifying aptamers that selectively target virtually any cell type under investigation. As an example, we provide the step-by-step protocol targeting CD19(+) Burkitt's lymphoma cells, starting from the pre-SELEX (systematic evolution of ligands by exponential amplification) measurements to establish suitable SELEX conditions and ending at completion of the SELEX procedure, which reveals the enriched single-stranded DNA library.

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 Dates: 2010
 Publication Status: Published in print
 Pages: -
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 Rev. Type: -
 Identifiers: Other: 21127492
DOI: 10.1038/nprot.2010.163
ISSN: 1750-2799 (Electronic)
ISSN: 1750-2799 (Linking)
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Title: Nature Protocols
  Alternative Title : Nat. Protoc.
Source Genre: Journal
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Pages: - Volume / Issue: 5 (12) Sequence Number: - Start / End Page: 1993 - 2004 Identifier: -