Fisheries and Aquatic Sciences

The Korean Society of Fisheries and Aquatic Science (한국수산과학회)
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A Versatile Method for DNA Sequencing of Unpurified PCR Products using an Automated DNA Sequencer and Tailed or Nested Primer Labeled with Near-infrared Dye: A Case Study on the Harmful Dinoflagellate Alexandrium

  • Ki Jang-Seu (Department of Life Science, College of Natural Sciences, Hanyang University) ;
  • Han Myung-Soo (Department of Life Science, College of Natural Sciences, Hanyang University)
  • Published : 2006.06.01
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Abstract

DNA sequence-based typing is considered a robust tool for the discrimination of dinoflagellate species because of the availability of extensive rDNA sequences. Here, we present a rapid, cost-effective DNA-sequencing technique for various PCR products. This sequencing strategy relies on 'nested' or 'tailed' primer labeled with near-infrared dye, and uses a minimal volume of unpurified PCR product (ca. $5{\mu}L$) as the DNA template for sequencing reactions. Reliable and accurate base identification was obtained for several hundred PCR fragments of rRNA genes. This quick, inexpensive technique is widely applicable to sequence-based typing in clinical applications, as well as to large-scale DNA sequencing of the same genomic regions from related species for studies of molecular evolution.

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References

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