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Updated: Jun 30, 2026
10:51
Fluorescence Based Primer Extension Technique to Determine Transcriptional Starting Points and Cleavage Sites of RNases In Vivo
Published on: October 31, 2014
DNA sequencing by synthesis with degenerate primers
Chao Tang1, Xiaolong Shi, Xiujie Li
1State Key Laboratory of Bioelectronics, Southeast University, Nanjing, China.
Journal of Genetics and Genomics = Yi Chuan Xue Bao
|September 23, 2008
Summary
Degenerate primer-based sequencing (DP-SBS) offers a reliable and cost-effective method for high-throughput DNA sequencing. This approach simplifies laboratory sequencing of short DNA fragments using conventional reagents.
Area of Science:
- Molecular Biology
- Genomics
- Biotechnology
Background:
- High-throughput DNA sequencing is crucial for genomic research.
- Existing sequencing methods can be complex and require specialized reagents.
Purpose of the Study:
- To develop and evaluate a novel degenerate primer-based sequencing by synthesis (DP-SBS) method.
- To assess the reliability, simplicity, and cost-effectiveness of DP-SBS for sequencing short DNA fragments.
Main Methods:
- Hybridization of degenerate primers to immobilized DNA templates on microarrays.
- Enzymatic primer extension by DNA polymerase with fluorescently labeled nucleotides.
- Detection of incorporated fluorescent nucleotides to determine template sequence (nine-base readout).
Main Results:
- DP-SBS accurately determined nine-base lengths of DNA sequences.
- The method utilizes conventional biochemical reagents, avoiding complex chemical treatments.
- DP-SBS demonstrated reliability and simplicity for laboratory-scale sequencing.
Conclusions:
- DP-SBS is a viable, cost-effective alternative for sequencing numerous short DNA fragments.
- The method's reliance on standard reagents simplifies laboratory implementation and reduces costs.
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