Whole-Genome Amplification From One Cell

A kit has been developed that supports whole-genome amplification (WGA) from a single cell, resulting in a million-fold amplification, yielding microgram quantities of genomic DNA. Nanogram quantities of DNA can be amplified up to 500-fold.

Genomic testing and characterization are important tools for understanding biologic systems and are dependent on the availability of sufficient quantities of genomic DNA. This is a problem for rare and archival sources of DNA. The GenomePlex WGA kits were produced by Sigma-Aldrich (St. Louis, MO, USA) for use as a high-throughput system for rapid and representative amplification of genomic DNA from trace amounts of starting material.

The new kit is derived from the proprietary amplification method that is based on random fragmentation of the genome into a series of overlapping, short templates. The resulting shorter DNA strands are efficiently primed and amplified to generate a library of DNA fragments with defined 3´ and 5´ termini, the OmniPlex library. This library is replicated using a linear, isothermal amplification in the initial stages, followed by a limited round of geometric polymerase chain reaction (PCR) amplifications.

Ninety-six samples can be prepared and amplified in under four hours. The automated method for GenomePlex is suitable for a variety of samples including buccal swabs, whole blood, blood cards, and formalin-fixed embedded tissues. It is accurate and eliminates human error and cross contamination. WGA is accomplished with minimal allelic drop out.

The kit will ease the concerns of limited patient material and provide sufficient DNA for downstream studies. It will advance cancer research, and aid genetic profiling and preimplantation genetic diagnostics.

The GenomePlex WGA technology is licensed to Rubicon Genomics (Ann Arbor, MI, USA). By combining GenomePlex WGA with related OmniPlex methods to amplify methylated DNA and total RNA, Rubicon has developed an integrated molecular diagnostic test for genetic, epigenetic, and expression factors.





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