ABI Prism® 373 and 377 DNA Sequencer
ABI offered two gel-based real-time instruments, the ABI Prism® 373 and 377. In the mid-to-late 1990’s, the ABI Prism® 377 DNA Sequencers were the most popular real-time instrument platforms for amplified DNA product separation. For the purpose of this discussion, the ABI Prism® 377 DNA Sequencers will be used, but it should be noted that both instrument platforms operate similarly.
The ABI Prism® 377 DNA Sequencers allow for simultaneous migration of amplified DNA products in different size ranges which are multiplexed with different color fluorophores. The ABI Prism® 373 and 377 DNA Sequencers, as well as the ABI capillary electrophoresis instruments allow for multi-component analysis which is the process that separates the different fluorescent dye colors into distinct spectral components. The process of fluorescence detection is discussed in the next module.
Summary
Slab-gel electrophoresis methods provided the forensic community with a mechanism to separate and detect amplified DNA. Today, with the advances in technology and the need for increased automation, the majority of the forensic community has moved to the capillary-based systems. However, a few laboratories still use the ABI Prism® 377 DNA Sequencer or FMBIO®.
CE offers ease of use, high resolution, minimal sample consumption, speed, on-line detection, minimized cross-contamination concerns, and is fully automatable. Additionally, CE provides quantitative information. The next module will cover CE methods in detail.
Journals presenting different electrophoretic techniques:
- Analytica Chimica Acta
- Analytical Biochemistry
- Chromatographia
- Electrophoresis
- Journal of Chromatography A
- Journal of Chromatography B
- Journal of Pharmaceutical and Biomedical Analysis
- Journal of Proteome Research
- Journal of Separation Science
- Proteomics
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