Many applications of electrophoresis involve separationof DNA fragments not only for analysis but also for sampleisolation and purification. Recovering DNA from electrophoreticgels and purifying it is an initial step in procedures such as thepolymerase chain reaction (PCR), developing cloning libraries forphysical mapping and DNA sequencing. Current rocedures forextracting DNA from a gel medium are tedious, produce inconsistentresults, and consume many hours of a skilled technical staff. These manually intensive procedures are clearly not suitable forprojects requiring high throughput such as the isolation ofhundreds of DNA fragments per day. In addition, present techniquesfor viewing DNA bands require staining with ethidium bromide dye,a potent inhibitor of many of the enzymes commonly used insubsequent cloning steps or in a PCR. This project will addressthese inadequacies by developing a semi-automated workstation toprovide direct visualization of electrophoretic patterns ofunstained DNA using electric birefringence as a detection scheme,machine vision techniques to locate and excise DNA bands, andautomated extraction of DNA from the gel matrix using speciallydesigned electroelution cells.Anticipated Results/Potential Commercial Applications as described by the awardee:Phase I will provide a vast improvement in thespeed and accuracy of recovering DNA bands from electrophoreticgels.
