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Related Experiment Videos

End sequence determination from large insert clones using energy transfer fluorescent primers

M Marra1, L A Weinstock, E R Mardis

  • 1Genome Sequencing Center, St. Louis, Missouri 63108, USA.

Genome Research
|November 1, 1996
PubMed
Summary
This summary is machine-generated.

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Obtaining high-quality DNA sequence data from large genomic clones is now feasible. This advancement aids in genome mapping, species comparison, and identifying overlapping DNA clones.

Area of Science:

  • Genomics
  • Molecular Biology
  • Bioinformatics

Background:

  • Genome mapping relies on confirming overlaps between DNA clones.
  • End-sequence data from large DNA inserts (>30 kb) can establish these overlaps.
  • Challenges include clone instability and the need for high DNA quantities for sequencing.

Purpose of the Study:

  • To develop and validate a method for routine, high-quality end-sequence data generation from large insert genomic clones.
  • To assess the feasibility of obtaining up to 800 bases of sequence data.

Main Methods:

  • Utilized ThermoSequenase DNA polymerase.
  • Employed Energy Transfer fluorescent primers for sequencing.
  • Focused on DNA derived from the ends of large insert genomic clones (>30 kb).

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Main Results:

  • Consistently obtained up to 800 bases of high-quality sequence data.
  • Demonstrated the feasibility of routine end-sequence data generation.
  • Validated the utility of this data for genome mapping applications.

Conclusions:

  • Routine, high-quality end-sequence data from large insert genomic clones is achievable.
  • This data is valuable for assessing clone overlaps and conservation of synteny.
  • Facilitates the detection of additional contiguous clones for comprehensive genome mapping.