High-level multiplex DNA amplification

We present data on efficient amplification of large number of DNA targets u sing a single-tube polymerase chain reaction (PCR). This is a further enhan cement of our approach to multiplexed PCR based on PCR suppression, which a llows multiple DNA amplification using only one sequence-specific primer pe r amplicon while the second primer is common for all targets (Broude, N.E., et al., Proc. Natl. Acad. Sci. USA 98, 206-211, 2001). The reaction condit ions have been optimized for simultaneous synthesis of 30 DNA targets, most ly consisting of fragments containing single nucleotide polymorphisms (SNP) . The size of the amplified fragments, derived from many different human ch romosomes, varies from 100 to 600 bp. We conclude that this method has pote ntial for highly multiplexed DNA amplification useful for SNP analyses, DNA diagnostics, and forensics.