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Updated: Aug 10, 2026
10:17
An Allele-specific Gene Expression Assay to Test the Functional Basis of Genetic Associations
Published on: November 3, 2010
A sensitive, quantitative assay for measurement of allele-specific transcripts differing by a single nucleotide
J Singer-Sam1, J M LeBon, A Dai
1Biology Division, Beckman Research Institute, City of Hope, Duarte, CA 91010.
PCR Methods and Applications
|February 1, 1992
Summary
The single nucleotide primer extension assay can now quantitatively measure allele-specific RNA transcripts. This PCR-based method accurately detects specific RNA even with a large excess of other RNA molecules.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- The single nucleotide primer extension assay is a polymerase chain reaction (PCR)-based technique.
- Currently, this assay is primarily used for qualitative measurement of allelic differences in DNA.
Purpose of the Study:
- To establish the quantitative application of the single nucleotide primer extension assay.
- To demonstrate its utility in measuring allele-specific RNA transcripts that differ by a single nucleotide.
Main Methods:
- Utilized the single nucleotide primer extension assay, a PCR-based method.
- Applied the assay to RNA samples for quantitative transcript analysis.
Main Results:
- The single nucleotide primer extension assay was successfully adapted for quantitative measurement.
- Demonstrated the ability to specifically detect the Pgk-1a transcript.
- Showed specific detection of Pgk-1a RNA even in the presence of a 1000-fold excess of Pgk-1b RNA.
Conclusions:
- The single nucleotide primer extension assay offers a novel quantitative approach for analyzing allele-specific transcripts.
- This method provides high specificity and sensitivity for detecting RNA molecules differing by a single nucleotide.
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