Validated Primer Collection for Mouse Gene Expression Analysis
Author Information
Author(s): Spandidos Athanasia, Wang Xiaowei, Wang Huajun, Dragnev Stefan, Thurber Tara, Seed Brian
Primary Institution: Center for Computational and Integrative Biology, Massachusetts General Hospital
Hypothesis
Can a comprehensive collection of experimentally validated primers improve the accuracy of transcript abundance measurement in mice?
Conclusion
The study successfully identified 17,483 validated primer pairs for murine transcripts that can be used under a common PCR thermal profile.
Supporting Evidence
- 17483 primer pairs were validated to amplify unique sequences corresponding to distinct murine transcripts.
- The primers were designed to work under a common PCR thermal profile.
- High-throughput validation procedures were employed to ensure primer specificity and efficiency.
- Successful amplification was confirmed through QPCR, gel electrophoresis, and sequencing.
Takeaway
The researchers created a big list of tested primers that help scientists measure how much of certain genes are present in mice, making it easier to study genes.
Methodology
The study involved designing and validating 26,855 primer pairs for mouse genes using QPCR, gel electrophoresis, and sequencing.
Limitations
Some primer pairs failed to amplify due to low sequence quality or absence of target sequences in the cDNA used.
Participant Demographics
The study focused on mouse genes, specifically using RNA from various mouse tissues.
Digital Object Identifier (DOI)
Want to read the original?
Access the complete publication on the publisher's website