Aryldiazomethanes for universal labeling of nucleic acids and analysis on DNA chips.
DNA and RNA labeling and detection are key steps in nucleic acid-based technologies, used in medical research and molecular diagnostics. We report here the synthesis, reactivity, and potential of a new type of labeling molecule, m-(N-Biotinoylamino)phenylmethyldiazomethane (m-BioPMDAM), that reacts selectively and efficiently with phosphates in nucleotide monomers, oligonucleotides, DNA, and RNA. This molecule contains a biotin as detectable unit and a diazomethyl function as reactive moiety. We demonstrate that this label fulfills the requirements of stability, solubility, reactivity, and selectivity for hybridization-based analysis and especially for detection on high-density DNA chips.
Laayoun
Ali
A
BioMérieux, Advanced Technology/Molecular Diagnostics, Marcy l'Etoile, France. ali.laayoun@eu.biomerieux.com
Kotera
Mitsuharu
M
Sothier
Isabelle
I
Trévisiol
Emmanuelle
E
Bernal-Méndez
Eloy
E
Bourget
Cécile
C
Menou
Lionel
L
Lhomme
Jean
J
Troesch
Alain
A
D019921:Biotinylation; D004247:DNA / Q000032:analysis* / Q000737:chemistry; D003978:Diazomethane / Q000031:analogs & derivatives* / Q000138:chemical synthesis; D005456:Fluorescent Dyes; D007202:Indicators and Reagents; D015394:Molecular Structure; D019906:Nuclear Magnetic Resonance, Biomolecular; D009693:Nucleic Acid Hybridization; D009696:Nucleic Acids / Q000032:analysis*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D012313:RNA / Q000032:analysis / Q000737:chemistry
D005456:Fluorescent Dyes; D007202:Indicators and Reagents; D009696:Nucleic Acids; D003978:Diazomethane; D012313:RNA; D004247:DNA