High-throughput sequencing of picogram amounts of DNA

CONECTUS ALSACE



15 Juillet 2015

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Fields

Biology / Medical

Sectors

Health

Versatile and robust DNA amplification method, for Genome-wide studies with picogram DNA amounts.

TECHNOLOGY

  • Single-tube RNA polymerase-based linear DNA amplification technology which facilitates global chromatin analyses with very limited cell populations, such as stem or cancer initiating cells.
  • Consecutive steps are performed in the same tube by sequential addition of reagents, thus eliminating the need for column purification and minimizing the risk of sample losses particularly at early steps.
  • Suitable for ChIP-seq and re-ChIP-seq profiling comparable to biological replicates.

APPLICATION

  • Genome-wide studies
  • Forensic use and archaeological sciences

INNOVATION ADVANTAGES

  • The single tube design of LinDA is ideal for handling ultra-small amounts of DNA (<30pg) and is compatible with automation
  • PCR-free procedure thus free of GC amplification bias
  • The actual hands-on working time is less than 6h with one overnight reaction
  • Can be coupled to a great number of global profiling technologies
  • Alows NGS data pre-processing thanks to the tool TrimLinDA

DEVELOPMENT STATUS

Detailed LinDA protocol describing all materials and critical steps, in addition to examples and controls available :

  1. Nature Methods 8, 565-567 (2011),
  2. Nature Protocols 7, 328–339 (2012),
  3. Protocol exchange (2012), DOI 10.1038/protex.2011.256

 

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