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Assaying the quality of cDNA libraries
F S Hagen1, C L Gray, J L Kuijper
1ZymoGenetics, Inc., Seattle, WA 98105.
Biotechniques
|April 1, 1988
Summary
Assessing cDNA library quality is crucial for research success. Probing with actin cDNA helps determine the percentage of actin-positive clones and clone completeness, improving the chances of finding full-length gene sequences.
Area of Science:
- Molecular Biology
- Genetics
- Biotechnology
Background:
- cDNA libraries are essential tools in molecular biology.
- Assessing the quality of a constructed cDNA library is critical for downstream applications.
- Actin, a ubiquitous housekeeping gene, serves as a reliable marker for library quality assessment.
Purpose of the Study:
- To evaluate the utility of actin cDNA probing for assessing cDNA library quality.
- To determine the correlation between actin-positive clone percentage and library completeness.
- To establish a method for predicting the success rate of isolating full-length clones of interest.
Main Methods:
- Construction of cDNA libraries.
- Probing of filter lifts and Southern blots using an actin cDNA probe.
- Quantification of actin-positive clones.
- Assessment of cDNA clone completeness.
Main Results:
- Probing with actin cDNA provides a reliable measure of library quality.
- The percentage of actin-positive clones directly correlates with library completeness.
- These quality metrics predict the likelihood of successfully isolating full-length clones.
Conclusions:
- Actin cDNA probing is an effective method for evaluating cDNA library quality.
- This assessment aids researchers in selecting high-quality libraries for gene cloning.
- Improved library quality assessment enhances the efficiency of molecular biology research.