This Article Full Text (PDF) References Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Add to Saved Citations Download to citation manager Request Permissions Request Reprints Add to My Marked Citations Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Citing Articles via Scopus Google Scholar Articles by Xuei, X. Articles by Warrior, U. Search for Related Content PubMed PubMed Citation Articles by Xuei, X. Articles by Warrior, U. Pubmed/NCBI databases Compound via MeSH Substance via MeSH Hazardous Substances DB BIOTIN Social Bookmarking                         What's this?

Use of SAM2® Biotin Capture Membrane in Microarrayed Compound Screening (µARCS) Format for Nucleic Acid Polymerization Assays

xxuei{at}iupui.edu

Caroline A. David

Tim R. Middleton

Ben Lim

Ron Pithawalla

Chih-Ming Chen

Rakesh L. Tripathi

David J. Burns

Usha Warrior

Global Pharmaceutical R&D, Abbott Laboratories, Abbott Park, IL.

Microarrayed compound screening format (µARCS) is a novel high-throughput screening technology that uses agarose matrices to integrate various biochemical or biological reagents in the assay. To evaluate the feasibility of using the µARCS technology for nucleic acid polymerization assays, the authors developed HIV reverse transcription (RT) and E1-dependent human papillomavirus (HPV) replication assays in this format. HIV RT is an RNA-dependent DNA polymerase, whereas HPV E1 is a DNA helicase. To ensure the efficient capture of the nucleic acid polymerization reaction and to minimize the nonspecific binding, the authors used a SAM²® biotin capture membrane in the assay. In both studies, the nucleic acid substrate was biotinylated on one end and was bound to the SAM²® membrane. A low melting-point agarose gel containing the rest of the reaction components was first placed on a polystyrene sheet spotted with compounds to allow passive diffusion of the compounds into the gel. The gel was removed from the compound sheet and applied to the SAM²® membrane with the immobilized nucleic acid template to initiate the polymerization. After the incubation, the membrane was washed with a high-salt buffer and exposed for imaging. Potential inhibitors can be seen as white spots on a dark background. The sensitivity for the known inhibitors appears to be comparable in µARCS as in a traditional 96-well plate assay. The methodology described in this paper further expands the applications of µARCS technology. (Journal of Biomolecular Screening 2003:273-282)

Key Words: µARCS • high-throughput screening • SAM2® • nucleic acid polymerization assays

Journal of Biomolecular Screening, Vol. 8, No. 3, 273-282 (2003)
DOI: 10.1177/1087057103008003005


CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

				X. Cheng, B. Yan, L. Gao, H. Tang, Y. Fan, S. N. Anderson, R. Affleck, and D. J. Burns Compound Transfer Efficiency from Polystyrene Surfaces: Application to Microarrayed Compound Screening J Biomol Screen, June 1, 2005; 10(4): 293 - 303. [Abstract] [PDF]

				D. R. Groebe, M. L. Maus, T. Pederson, J. Clampit, S. Djuric, J. Trevillyan, C. W. Lin, D. J. Burns, and U. Warrior Putting Thought to Paper: A {micro}ARCS Protease Screen J Biomol Screen, December 1, 2003; 8(6): 668 - 675. [Abstract] [PDF]