DIG Biotin Labeled Probe Protocols

Chem-Link Labeling of DNA or RNA with DIG or Biotin Protocol 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.

DIG Labeling of cDNA Probes 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.

DNA In Situ Hybridization with an Alkaline Phosphatase-Based Fluorescent Detection System 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.

Identification of Single Bacterial Cells using DIG-Labeled Oligonucleotides Protocol 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.


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

In Situ Hybridization to Human Metaphase Chromosomes using DIG-, Biotin- or Fluorochrome-Labeled DNA 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.

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

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

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

PCR Labeling of Ds DNA with the PCR DIG Probe Synthesis Kit or PCR Labeling Mixes 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.

Procedures for Labeling DNA, RNA, and Oligonucleotides with DIG, Biotin, or Fluorochromes 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.

Protocol for Nonradioactive In Situ Hybridization to Polytene Chromosomes with a DIG-labeled DNA 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.

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