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Highly instable (GATA)n-containing sequences of the mouse during the cloning process
R Studer1, C Kammerbauer, H Zischler
1Max-Planck-Institut für Psychiatrie, Martinsried, Germany.
Electrophoresis
|February 1, 1991
Summary
Researchers developed locus-specific probes for mouse DNA fingerprinting using (GATA)4 repeats. Cloning artifacts were observed, affecting repetitive and non-repetitive DNA sequences during the process.
Area of Science:
- Molecular Biology
- Genetics
Background:
- DNA fingerprinting is crucial for genetic analysis.
- Repetitive DNA sequences like (GATA)4 are valuable markers.
- Generating locus-specific probes can be challenging.
Purpose of the Study:
- To develop locus-specific probes for mouse DNA fingerprinting.
- To investigate alterations in DNA during the cloning process.
Main Methods:
- Genomic DNA digestion with HaeIII.
- In-gel hybridization with (GATA)4 oligonucleotide probe.
- Construction of a genomic library via direct subcloning.
- DNA sequence analysis.
Main Results:
- Successfully generated (GATA)4-positive subclones.
- Observed alterations in repetitive and adjacent non-repetitive DNA sequences.
- Identified both metastable and stable subclones.
- Detected changes in vector sequences.
Conclusions:
- The cloning process can introduce artifacts affecting DNA sequences.
- Understanding these artifacts is essential for accurate genetic analysis.
- Locus-specific probes were successfully isolated despite cloning-induced changes.