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Related Experiment Videos

Whole-genome multiple displacement amplification from single cells.

Claudia Spits1, Cédric Le Caignec, Martine De Rycke

  • 1Research Centre Reproduction and Genetics, Academisch Ziekenhuis, Vrije Universiteit Brussel, Brussels, 1090, Belgium.

Nature Protocols
|May 10, 2007
PubMed
Summary

Multiple displacement amplification (MDA) efficiently amplifies minute DNA quantities from single cells. This whole-genome amplification method yields high-molecular-weight DNA suitable for various downstream genetic analyses.

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Area of Science:

  • Molecular Biology
  • Genetics

Background:

  • Whole-genome amplification (WGA) is crucial for analyzing limited DNA samples.
  • Existing PCR-based WGA methods have limitations in DNA quality and coverage.

Purpose of the Study:

  • To detail a protocol for single-cell multiple displacement amplification (MDA).
  • To enable preimplantation genetic diagnosis and other applications using minute DNA quantities.

Main Methods:

  • Single-cell collection and lysis.
  • Multiple displacement amplification (MDA) using phi29 DNA polymerase.
  • Detailed step-by-step protocol for MDA reaction.

Main Results:

  • MDA generates high-molecular-weight DNA with superior genome coverage compared to PCR-based WGA.

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  • The protocol successfully amplifies DNA from single cells.
  • Yields 1-2 microg of DNA from a single cell in 3 hours.

    • Conclusions:

    • Single-cell MDA is an efficient method for whole-genome amplification.
    • The protocol is suitable for preimplantation genetic diagnosis, forensic science, and microdissected tissue analysis.
    • Amplified DNA supports diverse downstream applications including sequencing and genetic analysis.