Towards the development and characterization of a selective DNA purification protocol by capillary affinity gel electrophoresis.
Written in English
About the Edition
A purification protocol based on the selective capture of DNA targets from "real" samples was developed. The capture material was manufactured by the immobilization of DNA probes into polyacrylamide. The affinity gel was loaded inside capillaries, and samples of single-stranded DNA targets were introduced and eluted by electrophoresis. Characterization of the affinity gels was performed using fluorescently-labelled DNA probes and targets.The affinity gel selectively captured a complementary target, even in the presence of non-complementary and five-base pair mismatch targets. The yield of probe immobilization in the affinity gel was found to be 40%, with capture and elution efficiencies of 80-93% and 81-95%, respectively. Preliminary work on a fraction collection system to directly measure the amount of target eluted was also performed. Characterization of the performance of the affinity capture gel is important to allow for reproducible delivery of a purified sample for subsequent quantification of the target.
|Contributions||University of Toronto. DEpt. of Chemistry.|
|The Physical Object|
|Pagination||xiv, 155 leaves.|
|Number of Pages||155|
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