Byproduct with altered fluorescent properties is formed during standard deprotection step of hexachlorofluorescein labeled oligonucleotides
Bioconjugate Chemistry, ISSN: 1043-1802, Vol: 20, Issue: 8, Page: 1441-1443
2009
- 5Citations
- 10Captures
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Metrics Details
- Citations5
- Citation Indexes5
- CrossRef5
- Captures10
- Readers10
- 10
Article Description
HEX-labeled oligonucleotides obtained via typical synthetic protocols may contain more than 15% of material with altered spectral characteristics. We discovered hexachlorofluorescein residue transformation unknown earlier for standard DNA ammonolysis step. HEX residue reacts with ammonium hydroxide yielding acridine derivative, which has differed UV-VIS and fluorescent properties compared to HEX. Therefore, for critical bioassays where sensitivity and/or fluorescent signal differentiation (e.g., in quantitative or multiplexed assays) are essential, the careful RP-HPLC purification step is required. © 2009 American Chemical Society.
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