title: Self-seal reagent: evaporation control for molecular histology procedures without chambers, clips or fingernail polish.
abstract: Sensitive nucleic acid based detection methods such as in situ PCR, in situ RT-PCR and PRINS have great potential in the areas of developmental biology, pathogenesis and diagnostics. However, control of evaporation from in situ reactions is critical to ensure reliable data. Self-Seal Reagent, a component added directly to the in situ reaction mixture, effectively controls evaporation during in situ procedures by creating an evaporation-limiting barrier around the periphery of a standard cover glass as the reaction proceeds. At the end of the procedure, the cover glass is easily removed by soaking in an aqueous solution. A model is presented for how Self-Seal Reagent controls evaporation while maintaining reagent concentrations. Self-Seal Reagent is shown to be effective in the detection of HIV sequences in cells by in situ PCR.
authors.affiliation: MJ Research, Inc., Watertown, MA, USA.
authors.forename: D E
authors.initials: DE
authors.lastname: Sullivan
authors.forename: L E
authors.initials: LE
authors.lastname: Bobroski
authors.forename: O
authors.initials: O
authors.lastname: Bagasra
authors.forename: M
authors.initials: M
authors.lastname: Finney
mesh_terms: D003358:Cosmetics; D004279:DNA, Viral / Q000032:analysis; D004867:Equipment Design; D006678:HIV / Q000235:genetics; D006651:Histocytochemistry; D006801:Humans; D017403:In Situ Hybridization / Q000295:instrumentation / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods*; D013601:T-Lymphocytes / Q000821:virology; D014867:Water
chemical_list: D003358:Cosmetics; D004279:DNA, Viral; D014867:Water