Methods to increase nucleotide signals by raman scattering

C - Chemistry – Metallurgy – 12 – Q

Patent

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Details

C12Q 1/68 (2006.01) B01L 3/00 (2006.01) C07H 21/02 (2006.01) C12P 19/34 (2006.01) G01N 21/29 (2006.01)

Patent

CA 2478881

The methods and apparatus disclosed herein concern nucleic acid sequencing by enhanced Raman spectroscopy. In certain embodiments of the invention, nucleotides are covalently attached to Raman labels before incorporation into a nucleic acid (13). Exonuclease (15) treatment of the labeled nucleic acid (13) results in the release of labeled nucleotides (16, 130), which are detected by Raman spectroscopy. In alternative embodiments of the invention, nucleotides (16, 130) released from a nucleic acid (13) by exonuclease (15) treatment are covalently cross-linked to silver or gold nanoparticles (140) and detected by surface enhanced Raman spectroscopy (SERS), surface enhanced resonance Raman spectroscopy (SERRS) and/or coherent anti-Stokes Raman spectroscopy (CARS). Other embodiments of the invention concern apparatus (10, 100, 210) for nucleic acid sequencing.

L'invention porte sur des procédés et sur un appareil s'appliquant au séquençage de l'acide nucléique par spectroscopie Raman améliorée. Selon certaines formes d'exécution de l'invention, les nucléotides sont liés de manière covalente aux étiquettes Raman avant leur incorporation dans un acide nucléique (13). Le traitement à l'exonucléase (15) de l'acide nucléique marqué (13) entraîne la libération des nucléotides marqués (16, 130) qui sont détectés par spectroscopie Raman. Selon d'autres formes d'exécution de l'invention, les nucléotides (16, 130) libérés d'un acide nucléique (13) par un traitement à l'exonucléase (15) sont réticulés de manière covalente aux nanoparticules d'argent ou d'or (140) et sont détectés par spectroscopie Raman améliorée en surface (SERS), par spectroscopie Raman à résonance améliorée en surface (SERRS) et/ou par spectroscopie Raman anti-Stokes cohérente (CARS). Les autres formes d'exécution de cette invention sont des appareils (10, 100, 210) utilisés pour le séquençage de l'acide nucléique.

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