Detection of electrophore-labeled DNA and albumin by gas chromatography:  labile amide electrophoric release tags

Abdel-Baky, Samy; Allam, Kariman; Giese, Roger W.

doi:10.1021/ac00052a031

Cited by 5 publications

(2 citation statements)

References 7 publications

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“…In principle, the detection of electrophores by time-of-flight (TOF) EC-MS could be very sensitive and fast, helping to make it possible, with use of highly multiplexed electrophores, to sequence DNA quite rapidly. Some of this potential has been pointed out previously. − …”

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confidence: 79%

Electrophore Mass Tag Dideoxy DNA Sequencing

Bian²,

Wang³

et al. 1997

Anal. Chem.

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Toward a goal of dideoxy sequencing DNA utilizing electrophore labels, we prepared four electrophore-labeled DNA oligonucleotide primers. Each primer has a different electrophore and DNA sequence but a common glycol keto (alpha,beta-dihydroxyketo) release group. Cleavage of this latter group by either periodate oxidation or a thermal retroaldol reaction releases the electrophores for detection by mass spectrometry. Successful sequencing data with these primers was obtained by capillary electrophoresis on an ABI Model 310 after fluorescence dideoxy terminator cycle sequencing reactions were conducted. In a separate experiment, it was demonstrated that a cocktail of the four electrophore DNA primers could be detected as a dried sample spot by CO2 laser desorption/capillary collection/gas chromatography electron capture mass spectrometry. These results establish some feasibility for our long-term goal of high-speed multiplex electrophore mass tag dideoxy DNA sequencing. Ultimately we plan to use a higher number of electrophore mass tags and to rely on direct detection of the desorbed electrophores by electron capture time-of-flight mass spectrometry.

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confidence: 79%

Electrophore Mass Tag Dideoxy DNA Sequencing

Bian²,

Wang³

et al. 1997

Anal. Chem.

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“…Primers, synthesized with a 5′ C6 spacer and aminohexyl modification, were covalently conjugated by a photocleavable link to Masscode tags (Qiagen Masscode technology) (8,9). Masscode tags have a modular structure, including a tetrafluorophenyl ester for tag conjugation to primary amines; an o-nitrobenzyl photolabile linker for photoredox cleavage of the tag from the analyte; a mass spectrometry sensitivity enhancer, which improves the efficiency of atmospheric pressure chemical ionization of the cleaved tag; and a variable mass unit for variation of the cleaved tag mass (8,(10)(11)(12) primer sets are labeled with distinct molecular weight tags. Thus, amplification of a microbial gene target produces a dual signal that allows assessment of specificity.…”

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confidence: 99%