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Deoxynucleotides can replace dideoxynucleotides in minisequencing by arrayed primer extension.
Scott J Tebbutt1, Gareth D Mercer, Ron Do
1James Hogg iCAPTURE Centre for Cardiovascular and Pulmonary Research, St Paul's Hospital, University of British Columbia, Vancouver, Canada. stebbutt@mrl.ubc.ca
Biotechniques
|March 30, 2006
Summary
Array-based minisequencing can now use fluorescent deoxynucleotides, not just dideoxynucleotides. This finding offers a more cost-effective approach for DNA sequencing and genotyping applications.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- Array-based minisequencing technologies typically require fluorescent dideoxynucleotides for single-base extension.
- This method ensures accurate base calling and genotype determination through sequence-specific intensity data.
Purpose of the Study:
- To investigate the feasibility of using fluorescent deoxynucleotides in microarray minisequencing.
- To evaluate an alternative chemistry for array-based genotyping.
Main Methods:
- Experiments were conducted using fluorescent deoxynucleotides in microarray minisequencing.
- Fluorescent sequence extension intensity profiles were generated and analyzed.
Main Results:
- Fluorescent deoxynucleotides reliably produced intensity profiles comparable to those from dideoxynucleotides.
- The generated profiles were homologous to single-base extensions.
Conclusions:
- Fluorescent deoxynucleotides are a viable alternative to dideoxynucleotides in minisequencing.
- This approach offers a cost-effective and potentially IP-unencumbered method for array-based genotyping.