Methods to increase nucleotide signals by raman scattering
First Claim
1. A method comprising:
- a) obtaining nucleotides covalently linked to gold or silver, or gold and silver, nanoparticle(s), wherein the nucleotide and nanoparticle are linked via a terminal highly reactive cross-linking group selected from the group consisting of epoxide groups, azido groups, triazine groups, arylazido groups, and diazo groups;
b) synthesizing a nucleic acid comprising the labeled nucleotides;
c) immobilizing the nucleic acid of b) on a solid substrate;
d) sequentially releasing nucleotides from one end of the nucleic acid;
e) separating the released nucleotides from the immobilized nucleic acid by transferring the nucleotides through a microfluidic channel;
f) identifying nucleotides by Raman spectroscopy; and
g) determining the sequence of the nucleic acid.
1 Assignment
0 Petitions
Accused Products
Abstract
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.
108 Citations
7 Claims
-
1. A method comprising:
-
a) obtaining nucleotides covalently linked to gold or silver, or gold and silver, nanoparticle(s), wherein the nucleotide and nanoparticle are linked via a terminal highly reactive cross-linking group selected from the group consisting of epoxide groups, azido groups, triazine groups, arylazido groups, and diazo groups; b) synthesizing a nucleic acid comprising the labeled nucleotides; c) immobilizing the nucleic acid of b) on a solid substrate; d) sequentially releasing nucleotides from one end of the nucleic acid; e) separating the released nucleotides from the immobilized nucleic acid by transferring the nucleotides through a microfluidic channel; f) identifying nucleotides by Raman spectroscopy; and g) determining the sequence of the nucleic acid. - View Dependent Claims (2, 3, 4, 5, 6, 7)
-
Specification