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High Throughput MicroRNA Profiling: Optimized Multiplex qRT-PCR at Nanoliter Scale on the Fluidigm Dynamic ArrayTM IFCs
Published on: August 3, 2011
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A click chemistry-based microRNA maturation assay optimized for high-throughput screening.
Daniel A Lorenz1, Amanda L Garner
1Program in Chemical Biology, University of Michigan, 210 Washtenaw Avenue, Ann Arbor, Michigan 48109, USA.
Chemical Communications (Cambridge, England)
|June 11, 2016
Summary
Catalytic enzyme-linked click-chemistry assays (cat-ELCCA) now include the inverse-electron demand Diels-Alder (IEDDA) reaction. This advancement enhances assay sensitivity and reproducibility for automated high-throughput screening.
Area of Science:
- Biochemistry
- Chemical Biology
- Assay Development
Background:
- Catalytic enzyme-linked click-chemistry assays (cat-ELCCA) are a developing class of biochemical assays.
- Expanding the available reactions for cat-ELCCA can improve assay performance.
Purpose of the Study:
- To incorporate the inverse-electron demand Diels-Alder (IEDDA) reaction into the cat-ELCCA toolkit.
- To evaluate the impact of IEDDA on assay sensitivity and reproducibility.
Main Methods:
- Development of novel cat-ELCCA incorporating the IEDDA reaction.
- Testing and validation of the new assay format.
Main Results:
- Successful integration of the IEDDA reaction into cat-ELCCA.
- Demonstrated improvements in assay sensitivity and reproducibility compared to existing methods.
- Enabled automated high-throughput screening capabilities.
Conclusions:
- The IEDDA reaction expands the cat-ELCCA toolkit with superior kinetics.
- This enhanced cat-ELCCA technology offers improved performance for biochemical assays.
- The developed assay is suitable for automated high-throughput screening applications.
