Improved FISH Labelling Method for BAC Clones
Author Information
Author(s): Liesbeth Backx, Reinhilde Thoelen, Hilde Van Esch, Joris R. Vermeesch
Primary Institution: Center for Human Genetics, University Hospital Leuven
Hypothesis
The high background in traditional labelling methods for DOP amplified BAC clones can be reduced by using small degenerate oligonucleotide primers.
Conclusion
We developed a fast and accurate FISH labelling method that allows for the rapid generation of FISH probes without isolating BAC DNA.
Supporting Evidence
- The new method resulted in a 97% success rate for labelling BAC clones.
- Using small degenerate oligonucleotide primers reduced background signals in FISH.
- FISH can detect chromosomal aberrations that array CGH cannot.
Takeaway
We found a better way to label DNA samples so that scientists can see them more clearly under a microscope, making it easier to study chromosomes.
Methodology
We used degenerate oligonucleotide primers in a PCR process to label DOP amplified BAC clones for FISH analysis.
Limitations
FISH is not suitable for detecting small tandem duplications or deletions that are smaller than the probe used.
Digital Object Identifier (DOI)
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