Site-specific fluorescent labeling of DNA using Staudinger ligaton
Bioconjugate Chemistry, ISSN: 1043-1802, Vol: 14, Issue: 3, Page: 697-701
2003
- 73Citations
- 65Captures
- 1Mentions
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Example: if you select the 1-year option for an article published in 2019 and a metric category shows 90%, that means that the article or review is performing better than 90% of the other articles/reviews published in that journal in 2019. If you select the 3-year option for the same article published in 2019 and the metric category shows 90%, that means that the article or review is performing better than 90% of the other articles/reviews published in that journal in 2019, 2018 and 2017.
Citation Benchmarking is provided by Scopus and SciVal and is different from the metrics context provided by PlumX Metrics.
Metrics Details
- Citations73
- Citation Indexes73
- 73
- CrossRef54
- Captures65
- Readers65
- 65
- Mentions1
- References1
- 1
Article Description
We report the site-specific fluorescent labeling of DNA using Staudinger ligation with high efficiency and high selectivity. An oligonucleotide modified at its 5′ end by an azido group was selectively reacted with 5- [(N-(3′-diphenylphosphinyl-4′-methoxycarbonyl)phenylcarbonyl) aminoacetamido] fluorescein (Fam) under aqueous conditions to produce a Fam-labeled oligonucleotide with a high yield (∼90%). The fluorescent oligonucleotide was characterized by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Because of the relatively high yield of the Staudinger ligation, simple purification of the product by size-exclusion chromatography and desalting is sufficient for the resulting fluorescent oligonucleotide to be used as a primer in a Sanger dideoxy sequencing reaction to produce fluorescent DNA extension fragments, which are analyzed by a fluorescent electrophoresis DNA sequencer. The results indicate that the Staudinger ligation can be used successfully and site-specifically to prepare fluorescent oligonucleotides to produce DNA sequencing products, which are detected with single base resolution in a capillary electrophoresis DNA sequencer using laser-induced fluorescence detection.
Bibliographic Details
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