Related Experiment Videos
"Junction trapping". A simple PCR-based method for the isolation of YAC-insert termini
K Patel1, D Sheer, G M Hampton
1Human Cytogenetics Laboratory, Imperial Cancer Research Fund, London, England.
Genetic Analysis, Techniques and Applications
|April 1, 1993
Summary
A novel junction trapping technique efficiently isolates yeast artificial chromosome (YAC) insert termini. This PCR-based method reliably retrieves both ends of YAC inserts, proving highly effective in initial studies.
Area of Science:
- Molecular Biology
- Genomics
- Biotechnology
Background:
- Yeast artificial chromosomes (YACs) are crucial for cloning large DNA fragments.
- Efficient isolation of YAC insert termini is essential for genomic analysis and manipulation.
- Existing methods for termini isolation can be complex and time-consuming.
Purpose of the Study:
- To introduce a new, reliable method for isolating yeast artificial chromosome (YAC)-insert termini.
- To provide a streamlined approach for characterizing YAC ends.
Main Methods:
- The described method, "junction trapping," involves partial digestion of YAC-containing yeast strains.
- Ligation of digested DNA to a plasmid vector.
- Two rounds of polymerase chain reaction (PCR) are used for selection of terminal sequences.
Main Results:
- The junction trapping method successfully generated both terminal sequences from all nine examined YACs (100% success rate).
- The technique demonstrated high reliability and efficiency in isolating YAC insert termini.
Conclusions:
- Junction trapping is a potentially reliable and efficient new method for YAC insert termini isolation.
- This technique simplifies the characterization of YAC ends, advancing genomic research.