Abstract
Multiple displacement amplification (MDA) is a recently described method of whole-genome amplification (WGA) that has proven efficient in the amplification of small amounts of DNA, including DNA from single cells. Compared with PCR-based WGA methods, MDA generates DNA with a higher molecular weight and shows better genome coverage. This protocol was developed for preimplantation genetic diagnosis, and details a method for performing single-cell MDA using the φ29 DNA polymerase. It can also be useful for the amplification of other minute quantities of DNA, such as from forensic material or microdissected tissue. The protocol includes the collection and lysis of single cells, and all materials and steps involved in the MDA reaction. The whole procedure takes 3 h and generates 1–2 μg of DNA from a single cell, which is suitable for multiple downstream applications, such as sequencing, short tandem repeat analysis or array comparative genomic hybridization.
Note: In the version of this article initially published online, the third and fourth panels of Figure 1 (p. 1965) were incorrectly drawn. Figure 1 has been corrected in all versions of the article.
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Change history
21 December 2006
In the version of this article initially published online, the third and fourth panels of Figure 1 (p. 1965) were incorrectly drawn. Figure 1 has been corrected in all versions of the article.
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In the version of this article initially published online, the third and fourth panels of Figure 1 (p. 1965) were incorrectly drawn. Figure 1 has been corrected in all versions of the article.
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Spits, C., Le Caignec, C., De Rycke, M. et al. Whole-genome multiple displacement amplification from single cells. Nat Protoc 1, 1965–1970 (2006). https://doi.org/10.1038/nprot.2006.326
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DOI: https://doi.org/10.1038/nprot.2006.326
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