Universal fluorescent labeling (UFL) method for automated microsatellite analysis.

Masashi Shimizu; Natsuki Kosaka; Takashi Shimada; Takemitsu Nagahata; Hironori Iwasaki; Hisaki Nagai; Tadayoshi Shiba; Mitsuru Emi

doi:10.1093/dnares/9.5.173

PubMed · 12465717

Universal fluorescent labeling (UFL) method for automated microsatellite analysis.

Masashi Shimizu·Natsuki Kosaka·Takashi Shimada·Takemitsu Nagahata·Hironori Iwasaki·Hisaki Nagai·Tadayoshi Shiba·Mitsuru Emi

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Abstract

We have devised a novel method for automated microsatellite analysis using "universal" fluorescent labeling. This system is based on polymerase chain reactions driven by sequence-specific primers and a reporter primer labeled with a fluorescent dye at its 5' end. The forward sequence-specific primer is designed with a tag region bearing no homology to any human genomic sequence. Complementary tag sequences act as templates for the 6-carboxyfluorescein-labeled reporter primer, and those products can be analyzed with an autosequencer. The results we achieved with this assay system were consistent with the results of conventional assays using radioisotope-labeled primers, and diagnosis required less time. Furthermore, the fluorescent-labeled reporter primer is "universal" in that it can be used with different sequence-specific primers designed to carry the appropriate tag sequence at their 5'-ends. Our observations suggest that the "universal" fluorescent labeling method is an efficient tool for analyzing sequence variations in human DNA.

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Masashi Shimizu, Natsuki Kosaka, Takashi Shimada, Takemitsu Nagahata, Hironori Iwasaki, Hisaki Nagai, Tadayoshi Shiba, Mitsuru Emi. 2002-10-31. Universal fluorescent labeling (UFL) method for automated microsatellite analysis.. https://doi.org/10.1093/dnares%2F9.5.173

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