Molecular Encoding of Nucleic Acid Templates for Pcr and Sequence Analysis
Introduction The polymerase chain reaction (PCR) allows multiple copies of selected DNA sequences to be produced from limited amounts of a DNA template. Reactions with limited amounts of template, however, increase the risk of amplifying contaminant DNA and can also result in a skewed yield of PCR products such that there is a high degree of redundancy for a small portion of the original genomic sequences. Problems with contamination and redundancy are very common, particularly in PCR reactions with rare and irreplaceable DNA templates. Researchers at the UW have developed a simple and inexpensive method for verifying the authenticity of PCR products. Technology description Two novel methods for labeling PCR products have been developed. The first method uses an encoded oligonucleotide for Sequenase extension on a single target DNA strand prior to PCR amplification. The encoded oligonucleotide includes information specifying the date and the sample (batch stamp), a random nucleotide sequence (barcode), a region complementary to target locus, and a binding site for subsequent PCR amplification that is not complementary to any sequence in the genome. The second method incorporates the encoded information into hairpin-bisulfite PCR. This technique enables the analysis of epigenetic patterns on both the top and bottom strands of a DNA molecule simultaneously. The methods provide an authentication process for PCR reactions, distinguishing valid PCR products from contaminants and redundant sequences. Business opportunity While the technology has obvious applications in clinical PCR diagnostics and forensic testing, where accuracy of results is clearly a priority, the technology has the potential to become a universally adopted modification to the PCR method, particularly as it is relatively time and cost efficient. Stage of development A working model has been developed and research is ongoing to further validate and optimize the technique. Intellectual property position The UW has applied for patent protection to secure the rights to this technology.Type of Offer: Licensing
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