Labeling and detection of nucleic acids
View Patent ↗Provided in certain embodiments are new methods for forming azido modified nucleic acid conjugates of reporter molecules, carrier molecules or solid support. In other embodiments are provided methods for enzymatically labeling nucleic acids with an azide group.
1. A method of forming a nucleic acid conjugate, wherein the method comprises:
a) incorporating a terminal alkyne modified nucleotide into the nucleic acid polymer by contacting the terminal alkyne modified nucleotide with at least one other nucleotide in the presence of a DNA amplification enzyme to form a terminal alkyne modified nucleic acid polymer; and
b) contacting the terminal alkyne modified nucleic acid polymer with a carrier molecule or solid support that comprises an azido moiety to form a nucleic acid polymer-carrier molecule, or nucleic acid polymer-solid support conjugate.
2. The method according to claim 1 , wherein the carrier molecule is an amino acid, a peptide, a protein, a polysaccharide, a nucleotide, a nucleoside, an oligonucleotide, a nucleic acid, a hapten, a psoralen, a drug, a hormone, a lipid, a lipid assembly, a synthetic polymer, a polymeric microparticle, a biological cell or a virus.
3. The method according to claim 1 , wherein the carrier molecule comprises an antibody or fragment thereof, an avidin or streptavidin, a biotin, a blood component protein, a dextran, an enzyme, an enzyme inhibitor, a hormone, an IgG binding protein, a fluorescent protein, a growth factor, a lectin, a lipopolysaccharide, a microorganism, a metal binding protein, a metal chelating moiety, a non-biological microparticle, a peptide toxin, a phosphotidylserine-binding protein, a structural protein, a small-molecule drug, or a tyramide.
4. The method according to claim 1 , wherein the solid support is a microfluidic chip, a silicon chip, a microscope slide, a microplate well, silica gels, polymeric membranes, particles, derivatized plastic films, glass beads, cotton, plastic beads, alumina gels, polysaccharides, polyvinylchloride, polypropylene, polyethylene, nylon, latex bead, magnetic bead, paramagnetic bead, or superparamagnetic bead.
5. The method according to claim 1 , wherein the solid support is Sepharose, poly(acrylate), polystyrene, poly(acrylamide), polyol, agarose, agar, cellulose, dextran, starch, FICOLL, heparin, glycogen, amylopectin, mannan, inulin, nitrocellulose, diazocellulose or starch.
6. The method of claim 3 , wherein the non-biological microparticle is a nanoparticle.
7. The method of claim 4 , wherein the particle is a nanoparticle.
8. The method according to claim 1 , wherein the nucleic acid enzyme is a DNA polymerase.
9. The method according to claim 1 , wherein the nucleic acid enzyme is a RNA polymerase.