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Related Concept Videos

Labeling DNA Probes03:31

Labeling DNA Probes

DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
In-situ Hybridization02:31

In-situ Hybridization

In situ hybridization (ISH) is a technique used to detect and localize specific DNA or RNA molecules in cells, tissue, or tissue sections using a labeled probe. The technique was first used in 1969 for the investigation of nucleic acids. It is currently an essential tool in scientific research and clinical settings, especially for diagnostic purposes.
Types of probes and labels
A probe is a complementary strand of DNA or RNA that binds to corresponding nucleotide sequences in a cell. Many...
Southern Blot02:57

Southern Blot

Agarose gel electrophoresis is very useful in separating DNA fragments by size. Running a DNA ladder containing fragments of the known length alongside the sample helps determine the approximate length of the sample DNA fragments. However, additional steps are needed to verify the sequence identity of the sample DNA fragments.
Denatured DNA fragments must be transferred onto a carrier membrane from the gel to make it accessible to a probe - a small ssDNA fragment complementary to the target DNA...
FISH - Fluorescent In-situ Hybridization02:07

FISH - Fluorescent In-situ Hybridization

Fluorescence in situ hybridization, or FISH, was developed in the early 1980s and has quickly become one of the most widely used techniques in cytogenetics. Labeled probes are used to bind complementary DNA or RNA sequences on a chromosome or in a region within a cell. Earlier, the probes could only be obtained by cloning or reverse transcription of a DNA template. Currently, the probe oligonucleotides can be synthesized synthetically. Additionally, with the advancement of optical techniques,...

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Biotinylated probes in colony hybridization.

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Technical Demonstration of Whole Genome Array Comparative Genomic Hybridization
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Biotinylated probes in colony hybridization.

M J Haas1

  • 1U. S. Department of Agriculture, Eastern Reigonal Research Center, Philadelphia, PA.

Methods in Molecular Biology (Clifton, N.J.)
|November 15, 2012
PubMed
Summary

Colony hybridization detects specific DNA sequences in microbial colonies. This method uses a labeled probe to find and visualize target nucleic acids immobilized on a membrane.

Area of Science:

  • Molecular Biology
  • Microbiology

Background:

  • Colony hybridization is a key technique for identifying microbial colonies with specific genetic material.
  • Accurate detection of nucleic acid sequences is crucial in various biological and medical applications.

Purpose of the Study:

  • To describe the principle and methodology of colony hybridization for detecting nucleic acid sequences of interest.
  • To outline the steps involved in identifying positive hybridization events.

Main Methods:

  • Microbial colonies are grown and transferred to a membrane support.
  • Cell lysis and nucleic acid denaturation are performed on the membrane.
  • A labeled probe sequence is hybridized to the immobilized target nucleic acids.

Main Results:

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  • Successful hybridization indicates the presence of target nucleic acid sequences within the colonies.
  • The location of positive hybridization events can be determined on the membrane.

Conclusions:

  • Colony hybridization provides a reliable method for detecting specific nucleic acid sequences in microbial populations.
  • This technique facilitates the identification and isolation of cells containing desired genetic elements.