BioTechniques 31.- 1320-1324 (December 2001)

 

 

Increased Throughput for Fragment Analysis on an ABI PRISM® 377 Automated Sequencer Using a Membrane Comb and STRand Software

 

Robert J. Toonen and

Shayne Hughes

University of California, Davis, Davis, CA, USA

 

Abstract: This manuscript outlines our protocol for using a freely downloadable fragment analysis software package (STRand) togeth­er with a 96+4 RapidLoad membrane comb to increase throughput of samples for frag­ment analysis on ABI sequencers without costly upgrades from the manufacturer. We outline how using these products allows one to score 90 lanes of sample per gel on an ABI PRISM®377XL (64-lane sequencer), saving both time and money in the process­ing of samples. This protocol is a major modification to those suggested by the man­ufacturer. This protocol gives more consis­tent results that are easier to score than standard protocols, and it reduces reagent costs. Interest in fragment analysis (primar­ily microsatellites and AFLPs) is steadily increasing among both population ecolo­gists and geneticists, and methods that si­multaneously increase sample throughput while reducing costs associated with these analyses by over 50% per gel should prove useful to anyone using an ABI, MJ Base­stationTM, or LI-COR® automated se­quencer for fragment analysis.