Design of a fraction collector for capillary array electrophoresis.
This paper describes a prototype instrument for high-throughput fraction collection with capillary array electrophoresis (CAE). The design of the system was based on a comprehensive collection approach, in which fractions from all capillaries were simultaneously collected in individual collection microwells in predefined time intervals. The location of the fractions in the microwells on the collection plate was determined by monitoring the individual zone velocities close to the end of each capillary. The collection microwell plate was fabricated from buffer-saturated agarose gel, which maintained permanent electrical contact with the separation capillaries during the collection process. Since the collection gel plate consisted of over 90% water, liquid evaporation from the collection wells was minimized. A 12-capillary array instrument was built with two-point detection using a side illumination scheme. The collection performance was demonstrated by reinjection of selected fractions of a double-stranded DNA (dsDNA) separation. The identity of collected DNA fragments was confirmed by PCR and sequencing.
Minarik
Marek
M
Barnett Institute and Department of Chemistry, Northeastern University, Boston, MA 02115, USA.
Klepárnik
Karel
K
Gilár
Martin
M
Foret
Frantisek
F
Miller
Arthur W
AW
Sosic
Zoran
Z
Karger
Barry L
BL
D004247:DNA / Q000032:analysis / Q000302:isolation & purification*; D053938:DNA Fragmentation; D004262:DNA Restriction Enzymes / Q000378:metabolism; D019075:Electrophoresis, Capillary / Q000295:instrumentation*; D004867:Equipment Design; D005240:Feasibility Studies; D009838:Oligodeoxyribonucleotides / Q000032:analysis; D017422:Sequence Analysis, DNA
D009838:Oligodeoxyribonucleotides; D004247:DNA; D004262:DNA Restriction Enzymes