Michael Egholm. Molecular Staging, Inc.

Presenter - Dr. Seiyu Hosono

We have developed a method for effective single tube whole genome amplification of genomic DNA. The method, Multiple Displacement Amplification (MDA), faithfully replicates the gDNA with minimal loci and allele bias. Typically, 10 ng of DNA can be converted to 1 milligram of assay ready DNA in this simple isothermal reaction thus providing a ready resource of rare reference DNA. The basics of the technology have been described in two peer reviewed recent papers. (Proc. Nat. Acad. Sci., USA. 99(8), pp. 5261-5266 (2002); Gen. Res. 13:954-964 (2003)). The amplified DNA can be used directly, without any purification, in all standard nucleic acid based assays, including all PCR based methods. The reaction is scalable with a constant yield of 0.8 mg/mL independent of input of DNA and so far has been tested up to 10 mg yield of DNA. Furthermore, the MDA reaction can be used directly on crude cell lysates and hence the method allows for direct and simple isolation of DNA from complex matrices such as whole blood, lymphocytes, buccal swabs and blood spots.