Ultra-high throughput rotary capillary array electrophoresis scanner for fluorescent DNA sequencing and analysis.
We have constructed a rotary confocal fluorescence scanner and capillary array electrophoresis system that is designed to analyze over 1000 DNA sequencing or fragment sizing separations in parallel. Capillaries are arranged around the surface of a cylinder and a rotating objective in the middle of the cylinder excites and collects fluorescence from labeled DNA fragments as they pass the capillary detection window. The capillaries are pressure-filled with a replaceable matrix and the samples are electrokinetically injected in parallel from a stainless steel microtiter plate at the cathode end. We demonstrate that the instrument is capable of producing four-color data from all capillaries at a scan rate of 4 Hz (corresponding to a linear scan velocity of 121 cm/s). M13 sequencing data were obtained using a 128 capillary array mounted in half of the first quadrant of the scanner. In this initial run, read lengths greater than 500 bases were obtained in over 60% of the capillaries.
Scherer
J R
JR
Department of Chemistry, University of California, Berkeley 94720, USA.
Kheterpal
I
I
Radhakrishnan
A
A
Ja
W W
WW
Mathies
R A
RA
D017104:Bacteriophage M13 / Q000235:genetics; D001483:Base Sequence; D019075:Electrophoresis, Capillary / Q000295:instrumentation* / Q000379:methods*; D005456:Fluorescent Dyes / Q000032:analysis; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA / Q000295:instrumentation*; D012984:Software
D005456:Fluorescent Dyes