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Ultra-long Read Sequencing for Whole Genomic DNA Analysis
JoVE Journal
Bioengineering
This content is Free Access.
JoVE Journal Bioengineering
Ultra-long Read Sequencing for Whole Genomic DNA Analysis
DOI:

10:34 min

March 15, 2019

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Chapters

  • 00:04Title
  • 01:24High-molecular Weight (HMW) DNA Extraction
  • 03:11Ultra-long DNA Library Construction – Mechanical Shearing-based Protocol
  • 05:19Ultra-long DNA Library Construction – Transposase Fragmentation-based Protocol
  • 06:23Sequencing on the Nanopore Device
  • 07:52Results: Ultra-long DNA Sequencing Protocol
  • 09:38Conclusion

Summary

Automatic Translation

Long-read sequences greatly facilitate the assembly of complex genomes and characterization of structural variation. We describe a method to generate ultra-long sequences by nanopore-based sequencing platforms. The approach adopts an optimized DNA extraction followed by modified library preparations to generate hundreds of kilobase reads with moderate coverage from human cells.

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