Related Experiment Videos
Efficiency and specificity of gene isolation by exon amplification
M A North1, P Sanseau, A J Buckler
1Genome Analysis Laboratory, Imperial Cancer Research Fund, London, UK.
Summary
Exon amplification effectively identifies transcribed sequences from cosmids, successfully isolating exons from known genes with high efficiency. This method aids in gene discovery and characterization, complementing other genomic strategies.
Area of Science:
- Genomics
- Molecular Biology
- Gene Discovery
Background:
- Exon amplification is a valuable technique for identifying transcribed sequences.
- It complements existing strategies for gene isolation.
- Cosmids are frequently used to study complex genomic regions.
Purpose of the Study:
- To evaluate the efficiency, specificity, and reproducibility of exon amplification.
- To assess artefact formation during routine cosmid screening.
- To characterize novel exons cloned from human genomic DNA.
Main Methods:
- Exon amplification was performed on 32 cosmids, including 8 from the human MHC class II region on Chromosome 6.
- Efficiency was tested on known genes, including those with differential splicing.
- Artefact amplifications were analyzed for their nature and frequency.
Main Results:
- At least one exon was cloned from 88% of known genes tested.
- Artefact amplifications occurred in 19% of cosmids, often from noncoding sequences with splice junction homology.
- The procedure proved successful across various cosmid types and gene structures.
Conclusions:
- Exon amplification is an efficient and reproducible method for identifying transcribed exons.
- The technique is adaptable to diverse cosmid libraries and gene architectures.
- This approach facilitates the characterization of novel exons and gene discovery.