Application of high-resolution capillary array electrophoresis with automated fraction collection for GeneCalling™ analysis of the yeast genomic DNA

Capillary array instrument was applied to transcript profiling of the yeast genomic DNA using GeneCalling™ chemistry. The instrument integrated a 12‐capillary array for DNA separation with a replaceable sieving matrix, laser‐induced fluorescence detection and an automated microfraction collector. Th...

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Veröffentlicht in:Electrophoresis 2003-02, Vol.24 (4), p.639-647
Hauptverfasser: Berka, Jan, Ruiz-Martinez, Marie C., Hammond, Richard, Minarik, Marek, Foret, Frantisek, Sosic, Zoran, Kleparnik, Karel, Karger, Barry L.
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container_end_page 647
container_issue 4
container_start_page 639
container_title Electrophoresis
container_volume 24
creator Berka, Jan
Ruiz-Martinez, Marie C.
Hammond, Richard
Minarik, Marek
Foret, Frantisek
Sosic, Zoran
Kleparnik, Karel
Karger, Barry L.
description Capillary array instrument was applied to transcript profiling of the yeast genomic DNA using GeneCalling™ chemistry. The instrument integrated a 12‐capillary array for DNA separation with a replaceable sieving matrix, laser‐induced fluorescence detection and an automated microfraction collector. The DNA fractions, exiting the separation capillaries, were continuously deposited in a 1536‐well collection plate made of agarose gel. DNA fragments recovered from selected fractions were cloned and then sequenced. Over 80% of theoretically predicted fragments could be recovered in the collected fractions, cloned and sequenced with an average redundancy of threefold. Excellent correlation of the experimentally obtained sequences with the theoretically predicted gene fragments demonstrated the suitability of capillary array electrophoresis for micropreparative recovery of DNA fragments. This approach, useful especially for rapid DNA expression profiling of unknown genes for nonsequenced organisms, demonstrates the practical capability of the prototype multicapillary fraction collector.
doi_str_mv 10.1002/elps.200390075
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subjects Capillary array
Fraction collection
Gene expression profiling
title Application of high-resolution capillary array electrophoresis with automated fraction collection for GeneCalling™ analysis of the yeast genomic DNA
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