Screening cDNA Libraries for Full-Length Coding Sequences
Author Information
Author(s): Jacob Pollier, Miguel González-Guzmán, Wilson Ardiles-Diaz, Danny Geelen, Alain Goossens
Primary Institution: Vlaams Instituut voor Biotechnologie (VIB), Ghent University, Belgium
Hypothesis
Can a medium-throughput screening strategy effectively identify full-length coding sequences from incomplete cDNA tags?
Conclusion
The developed strategy efficiently converts incomplete expressed sequence tags to full-length coding sequences.
Supporting Evidence
- The method identified 109 full-length clones from 163 cDNA-AFLP tags, achieving a success rate of 67%.
- Multiple probes can be used in a single hybridization event, increasing throughput.
- The strategy allows for the screening of approximately 100,000 cDNA clones in a rapid manner.
Takeaway
The researchers found a way to quickly identify complete gene sequences from partial ones, which helps in studying plant genes better.
Methodology
A medium-throughput screening strategy combining PCR and colony hybridization was used to identify full-length clones from cDNA libraries.
Limitations
The method may not be specific for full-length clones and relies on the quality of the cDNA library.
Digital Object Identifier (DOI)
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