Increased throughput for fragment analysis on an ABI PRISM® 377 automated sequencer using a membrane comb and STRand software

• Published in: BioTechniques Vol. 31; no. 6; pp. 1320 - 1324
• Main Authors: TOONEN, Robert J, HUGHES, Shayne
• Format: Journal Article
• Language: English
• Published: Natick, MA Eaton 01.12.2001
• Subjects: Biological and medical sciences; Computational Biology; Diverse techniques; Fundamental and applied biological sciences. Psychology; Microsatellite Repeats - genetics; Molecular and cellular biology; Polymerase Chain Reaction; Polymorphism, Genetic - genetics; Sequence Analysis, DNA - instrumentation; Sequence Analysis, DNA - methods; Software; Sequencer; Automation; Biochemical analysis; Method; Fragment; Abies; Gymnospermae; Membrane; Coniferales; Spermatophyta; Software; Automatic analysis; Strand
• ISSN: 0736-6205; 1940-9818

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 Basestation, or LI-COR automated sequencer for fragment analysis.