RNA-RNA in situ hybridization using digoxigenin-labeled probes: the use of high-molecular-weight polyvinyl alcohol in the alkaline phosphatase indoxyl-nitroblue tetrazolium reaction.
A nonradioactive RNA-RNA in situ hybridization method using digoxigenin-labeled probes is described. The visualization of hybrids is done using the indoxyl-nitro blue tetrazolium alkaline phosphatase reaction. The addition of polyvinyl alcohols of high molecular weight (40-100 kDa) to the reaction medium enhances the alkaline phosphatase reaction and prevents diffusion of reaction intermediates, resulting in a 20-fold increase in sensitivity without increasing the background. Due to the more localized precipitation of the formazan, the site of hybridization can be determined more precisely.
De Block
M
M
Plant Genetic Systems N.V., Ghent, Belgium.
Debrouwer
D
D
D000107:Acetylation; D000469:Alkaline Phosphatase; D004076:Digoxigenin; D017403:In Situ Hybridization / Q000379:methods*; D007211:Indoles; D009580:Nitroblue Tetrazolium; D011142:Polyvinyl Alcohol; D012313:RNA / Q000032:analysis*; D015347:RNA Probes
D007211:Indoles; D015347:RNA Probes; D009580:Nitroblue Tetrazolium; C035455:5-bromo-4-chloro-3-indoxyl phosphate; D012313:RNA; D011142:Polyvinyl Alcohol; D000469:Alkaline Phosphatase; D004076:Digoxigenin