Transgenomic, Inc.

Kimberly A Lamb

Douglas T Gjerde

Paul D Taylor

Robert M Haefele

Christopher P Hanna

Satyajit Kar

Laura Bunnel

Jeffrey L Sklar

An apparatus for effecting base pair length separations of DNA fragments by matched ion paired chromatography comprising a separation column containing separation media having non-polar DNA separation surfaces, separation solution supply means, and a separation solution conduit communicating with the separation column and the separation solution supply means, and a cleaning solution valve means positioned in the separation solution conduit for injecting cleaning solution into the separation solution conduit. A process for cleaning the non-polar DNA separation surfaces in the apparatus comprising interrupting the flow of separation solvent with a block of cleaning solution injected into the flow of separation solution passing to the column, the cleaning solution containing agent which removes accumulated residues from the non-polar surface. The cleaning solution can have an alkaline pH and contain a chelating agent such as EDTA.

MIPC column cleaning system and process

A method for detecting a putative mutant DNA in a sample of DNA includes the steps of amplifying the sample of DNA using PCR; hybridizing the amplified sample to form a mixture of homoduplexes and heteroduplexes; separating the mixture into fractions by Denaturing Matched Ion Polynucleotide Chromatography; and blind collecting the eluted fractions at a retention time corresponding to the retention time of the heteroduplex. The DNA in the blind collected fractions can be PCR amplified to obtain an increased amount of heteroduplex relative to homoduplex. The method is useful for determining the remission status of a patient in which the tissue-derived DNA sample contains a large background of wild type or where the putative mutant DNA is below the limit of detection.

Target DNA amplification by MIPC and PCR

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Omaha, NE, United States of America

Kimberly A Lamb