Digoxigenin Labelled Probes, Non Radioactive Labelling Probes, Anti Dioxigenin, Alkaline Phosphate, Hybridization, Anti Digoxigenin Fluorescein

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Digoxigenin Labelled Probes

Digoxigenin is another chemical derived from plants and used for non-radioactive labelling of probes. An antibody associated with an enzyme (anti-digoxigenin-alkaline-phosphatase conjugate) is used for the detection of the presence of digoxigenin.

The probe may be labelled with digoxigenin-ll-dUTP supplied with a digoxigenin kit (these kits are available from any commercial firm, e.g. Boehringer Mannheim). The labelled and denatured probe may be used for hybridization with denatured DNA on Southern blots or for in-situ hybridization.

After hybridization the membrane or the slide may be washed and transferred into detection buffer containing 20 μg/ml of anti-digoxigenin fluorescein and 5% (w/v) BSA (bovine serum albumin). The system is incubated for one hour at 3TC, and then the membrane/slide is washed in detection buffer three times (8 min each at 3TC). Alkaline phosphatase activity can be detected using 0.17 mg/ml BCIP (5-bromo-4chloro-3 indolyl phosphate) and 0.33 mg/ml NBL (nitroblue tetrazolium) as dye substrate.

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c DNA Probes Synthetic Oligonucleotides as Probes RNA Probes or Riboprobes Labelling of Probes Radiolablled Probes Non Radioactive Probes Biotin Labelled Probes Digoxigenin Labelled Probes Alternatives to Biotin and Digoxigenin Labelling Amplification of DNA Probe Signals Techniques Used in Molecular Probing Southern Nothern and Western Blotting Southern Blotting Northern Blotting Western Blotting Dot Blots and Slot Blots Applications of Molecular Probes Construction and Screening of Genomic and cDNA Libraries Genomic Library by Shotgun Experiment cDNA Library from mRNAs Colony or Plaque Hybridization for Screening of Libraries Chromosome Walking and Characterization of Chromosome Segments Chromosome Jumping or Hopping Libraries BAC Libraries and Assembly of BACs into Contigs