How the ABI Prism 310 Genetic Analyzer Works

The ABI PRISM® 310 Genetic Analyzer is a system composed of instrument
hardware,
a Macintosh® computer, several types of software, and consumables.
This overview will help you identify parts of the ABI PRISM® 310 Genetic Analyzer
and
understand how it works.

DNA sequencing experiments determine the order of the bases in a DNA sample.
Fluorescently labeled dyes are attached to ACGT extension products in DNA
sequencing reactions. Dye labels are incorporated using either 5´-dye labeled
primers
or

3´-dye

labeled

dideoxynucleotide

terminators.

Polymerases

such

as

AmpliTaq® FS
are used for primer extension.
The sequencing reaction sample
tubes are placed in a tray in the
instrument’s
autosampler. The autosampler
successively brings each sample
into contact with the
cathode electrode and one end
of a glass capillary filled with
polymer. An anode
electrode at the other end of the
capillary is immersed in buffer.
A portion of the sample enters
the capillary as current flows
from the cathode to the
anode. This is called electrokinetic injection. The end of the capillary near the
cathode
is then placed in buffer. Current is applied again to continue electrophoresis.
When the nucleotides reach a detector window in the capillary coating, a laser
excites
the fluorescent dye labels. Emitted fluorescence from the dyes is collected by a CCD
camera. The software interprets the result, calling the bases from the fluorescence
intensity at each data point.