Detection of Digoxigenin-Labeled DNA Probes Hybridized to Plant Chromosomes In Situ
Digoxigenin is widely used as a nonradioactive label for in situ hybridization to locate DNA sequences along chromosomes in plants (e.g., 1, 2, 3) and animals. In many cases, its use is similar to biotin ( chapter 25, or e.g., 4), but digoxigenin labels may give lower unspecific background signal and therefore enable more efficient detection of short or low copy sequences. Digoxigenin is usually incorporated into the DNA enzymatically by nick translation, random priming, polymerase chain reaction (PCR), or end labeling using the modified nucleotide digoxigenin-11-dUTP (see chapter 10 and chapter 11).
KeywordsHybridization Signal Hydrogen Peroxide Solution Detection Buffer Tungstosilicic Acid Posthybridization Wash
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