New dye-labeled terminators for improved DNA sequencing patterns

• Published in: Nucleic acids research 1997-11, Vol.25 (22), p.4500-4504
• Main Authors: Rosenblum, B.B, Lee, L.G, Spurgeon, S.L, Khan, S.H, Menchen, S.M, Heiner, C.R, Chen, S.M
• Format: Article
• Language: English
• Subjects: 4'-aminomethylfluorescein; 4,7-dichlororhodamine; 5-carboxy-d-rhodamine; automation; bigdye terminators; chemical reactions; dichlororhodamine terminators; dna sequencing termination; DNA, Bacterial - analysis; DNA, Bacterial - chemistry; dyes; Energy Transfer; fluorescein; Fluoresceins - chemistry; fluorescent dyes; Fluorescent Dyes - chemistry; isomers; Molecular Structure; nucleotides; Rhodamines - chemistry; sequence analysis; Sequence Analysis, DNA - methods
• ISSN: 0305-1048; 1362-4962; 1362-4962
• DOI: 10.1093/nar/25.22.4500

We have used two new dye sets for automated dye-labeled terminator DNA sequencing. One set consists of four, 4,7-dichlororhodamine dyes (d-rhodamines). The second set consists of energy-transfer dyes that use the 5-carboxy-d-rhodamine dyes as acceptor dyes and the 5- or 6-carboxy isomers of 4′-aminomethylfluorescein as the donor dye. Both dye sets utilize a new linker between the dye and the nucleotide, and both provide more even peak heights in terminator sequencing than the dye-terminators consisting of unsubstituted rhodamine dyes. The unsubstituted rhodamine terminators produced electropherograms in which weak G peaks are observed after A peaks and occasionally C peaks. The number of weak G peaks has been reduced or eliminated with the new dye terminators. The improved signal-to-noise ratio of the energy-transfer dye-labeled terminators combined with more even peak heights results in successful sequencing of high molecular weight DNA templates such as bacterial artificial chromosome DNA.