T. Irie et al., Automated DNA fragment collection by capillary array gel electrophoresis array gel electrophoresis in search of differentially expressed genes, ELECTROPHOR, 21(2), 2000, pp. 367-374
An automatic DNA fragment collector using capillary array gel electrophores
is has been developed. A sheath flow technique is used for not only detecti
on but also collection of DNA fragments. In a sheath flow cell, the DNA fra
gments separated by 16 capillaries flow independently into corresponding sa
mpling capillaries. The fraction collector consists of 16 sampling trays an
d each sampling tray is set beneath each end of the sampling capillaries to
collect the flow-through DNA fragments. Certain DNA fragments are automati
cally sorted by controlling the movement of the sampling trays according to
the signals from the system. The collector experimentally separated two mi
xtures of polymerase chain reaction (PCR) products: one prepared by using e
ight different sizes (base lengths from 161 to 562) of DNAs; and the other
prepared by a differential display (DD) method with cDNA fragments. Collect
ed DNA fragments are amplified by PCR and measured by electrophoresis. DNA
fragments with base length differences of one (base lengths 363 and 364) we
re successfully separated. A separated DNA fragment from the DD sample was
also successfully sequenced. In addition, differentially expressed DNA frag
ments were automatically sorted by comparative analysis, in which two simil
ar cDNA fragment groups, labeled by two different fluorophores, respectivel
y, were analyzed in the same gel-filled capillary. These results show that
the automatic DNA fragment collector is useful for gene hunting in research
fields such as drug discovery and DNA diagnostics.