1. The ABI Prism 310 Genetic Analyzer
Chapter 11
The ABI Prism 310 Genetic Analyzer
©2002 Academic Press

2. ABI Prism 310 Genetic Analyzer
capillary
Syringe with polymer solution
Autosampler tray
Outlet buffer
Injection electrode
Inlet buffer
©2002 Academic Press

3. Close-up of ABI Prism 310 Sample Loading Area
Autosampler Tray
Sample Vials
Electrode
Capillary
©2002 Academic Press

4. Sample Processing Steps with ABI 310
Replace capillary
Refill syringe with polymer solution
Fill buffer vials
Performed only once per batch of ~96 samples
Prepare samples
(denature, cool, and mix with size standard)
Prepare sample sheet and injection list
Allelic ladder every tenth injection
©2002 Academic Press

5. Sample Processing Steps (cont.)
Automated Sample Injection, Electrophoresis and Data Collection
Size DNA Fragments
GeneScan Software
Genotype STR alleles
Genotyper Software
Perform Data Analysis
Manually inspect the data
ELECTROPHORESIS and DETECTION steps are simultaneous
©2002 Academic Press

6. Quality of Formamide Affects Sensitivity
©2002 Academic Press
Figure courtesy of Bruce McCord, Ohio University

7. Steps Performed in Standard Module
Capillary fill – polymer solution is forced into the capillary by applying a force to the syringe 
Pre-electrophoresis – the separation voltage is raised to 10,000 volts and run for 5 minutes;
Water wash of capillary – capillary is dipped several times in deionized water to remove buffer salts that would interfere with the injection process 
Sample injection – the autosampler moves to position A1 (or the next sample in the sample set) and is moved up onto the capillary to perform the injection; a voltage is applied to the sample and a few nanoliters of sample are pulled onto the end of the capillary; the default injection is 15 kV (kilovolts) for 5 seconds
Water wash of capillary – capillary is dipped several times in waste water to remove any contaminating solution adhering to the outside of the capillary
Water dip – capillary is dipped in clean water (position 2) several times
Electrophoresis – autosampler moves to inlet buffer vial (position 1) and separation voltage is applied across the capillary; the injected DNA molecules begin separating through the POP-4 polymer solution
Detection – data collection begins; raw data is collected with no spectral deconvolution of the different dye colors; the matrix is applied during Genescan analysis
©2002 Academic Press