DIG Biotin Labeled Probe Protocols

This protocol describes how to use DIG Chem-Link to directly label any DNA [e.g. plasmids, PCR products, cDNA prepared from mRNA] or RNA (e.g. total RNA, poly(A)+ mMRNA). The DIG Chem-Link or Biotin Chem-Link may also be used to label oligonucleotides. Includes: Required Purity of DIG Chem-Link Templates; Direct DIG Labeling of mRNA or cDNA with DIG Chem-Link; Key Product Required for Direct Labeling of DNA or RNA; Estimating the Yield of DIG-labeled Nucleic Acids. - [Read Chem-Link Labeling of DNA or RNA with DIG or Biotin Protocol]

Protocol for DIG labeling of cDNA probes. Includes: Incorporation DIG during PCR; Estimating the yield of DIG labeled probes; DNA Dot Blotting; Prehybridization and Hybridization. - [Read DIG Labeling of cDNA Probes Protocol]

Protocol describes here a high sensitivity indirect detection procedure for DIG-labeled hybridization probes. The procedure uses the components of the HNPP Fluorescent Detection Set to form a fluorescent precipitate of HNPP (2-hydroxy-3-naphthoic acid-2’-phenylanilide phosphate) and Fast Red TR at the site of hybridization. Includes: In situ hybridization with DIG-labeled probes; Detection of DIG-labeled probes; Fluorescence microscopy. - [Read DNA In Situ Hybridization with an Alkaline Phosphatase-Based Fluorescent Detection System]

Protocol for the identification of single bacterial cells using DIG-labeled oligonucleotides. Includes: Organisms and growth conditions; Cell fixation and preparation of cell smears; DIG labeling of oligonucleotides with DIG-ddUTP; In situ hybridization using digoxigenin-labeled oligonucleotides; Detection of DIG-labeled oligonucleotides with fluorescently labeled anti-DIG Fab fragments; Detection of DIG-labeled oligonucleotide.
 - [Read Identification of Single Bacterial Cells using DIG-Labeled Oligonucleotides Protocol]

In situ hybridisation to alpha satellite sequences (chromosome specific) protocol. - [Read In Situ Hybridisation to Alpha Satellite Sequences (Chromosome Specific) Protocol]

Protocol for In situ hybridization to human metaphase chromosomes using DIG-, biotin-, or fluorochrome-labeled DNA probes and detection with fluorochrome conjugates. Includes: Pretreatment of metaphase spreads on slides; Denaturation and hybridization; Single color fluorescent detection with immunological amplification; Multicolor fluorescence in situ hybridization (Multicolor FISH); Results obtained with human metaphase chromosome spreads. - [Read In Situ Hybridization to Human Metaphase Chromosomes using DIG-, Biotin- or Fluorochrome-Labeled DNA]

Protocol for nick-translation labeling of ds DNA with DIG-, Biotin-, or Fluorochrome-labeled dUTP.
- [Read Nick Translation Labeling of Ds DNA with DIG-, Biotin-, or Flurochrome- Labeled dUTP Protocol]

Protocol for oligonucleotide 3’-end labeling with DIG-ddUTP or Biotin-ddUTP.
- [Read Oligonucleotide 3’-end Labeling with DIG-ddUTP or Biotin-ddUTP Protocol]

Protocol for oligonucleotide tailing with a DIG-dUTP, Biotin-dUTP, or Fluorescein-dUTP mixture.
- [Read Oligonucleotide Tailing with a DIG-dUTP, Biotin-dUTP, or Fluorescein-dUTP Mixture Protocol]

Protocol for PCR labeling of ds DNA with the PCR DIG Probe Synthesis Kit or PCR Labeling Mixes. Includes: PCR DIG labeling reaction for moderately labeled probes; PCR fluorescein labeling reaction for direct in situ probes.
- [Read PCR Labeling of Ds DNA with the PCR DIG Probe Synthesis Kit or PCR Labeling Mixes]

Procedures for labeling DNA, RNA, and oligonucleotides with DIG, biotin, or fluorochromes. Includes: Random primed labeling of ds DNA with DIG-, Biotin- or Fluorescein-High Prime reaction mix; PCR labeling of ds DNA with the PCR DIG Probe Synthesis Kit or PCR Labeling Mixes. - [Read Procedures for Labeling DNA, RNA, and Oligonucleotides with DIG, Biotin, or Fluorochromes]

The protocol given makes the method of in situ hybridization easier, faster, more reliable, and available to anyone who can operate a microscope. Includes: Labeling the hybridization probe; Preparation and denaturation of polytene chromosomes from Drosophila, Chironomus, or other species; Hybridization and detection. - [Read Protocol for Nonradioactive In Situ Hybridization to Polytene Chromosomes with a DIG-labeled DNA]

Protocol for RNA labeling by in vitro transcription of DNA with DIG, Biotin or Fluorescein RNA Labeling Mix. - [Read RNA Labeling by In Vitro Transcription of DNA with DIG, Biotin or Fluorescein RNA Labeling Mix]