Mass spectrometric methods for detecting mutations in a target nucleic acid
View Patent ↗Provided herein are mass spectrometric methods for detecting a mutation in a target nucleic acid. In an embodiment, provided is a method which comprises digesting a target nucleic acid molecule; capturing digested fragments on a solid support that comprises oligonucleotides complementary thereto; and detecting hybrids and the molecular weights of captured fragments by mass spectrometry, thereby identifying mutations in the target nucleic acid molecule.
1. A method for detecting mutations in a target nucleic acid, comprising: digesting a target nucleic acid molecule; capturing digested fragments on a solid support that comprises oligonucleotides complementary thereto; detecting hybrids and the molecular weights of captured fragments by mass spectrometry, thereby identifying mutations in the target nucleic acid molecule.
2. The method of claim 1 , wherein the target molecule is RNA.
3. The method of claim 2 , wherein the RNA is produced by transcription of a target DNA molecule.
4. The method of claim 1 , wherein the solid support comprises a flat surface.
5. The method of claim 1 , wherein the solid support comprises a matrix for performing matrix-assisted laser desorption/ionization (MALDI) mass spectrometry.
6. The method of claim 1 , wherein the solid support is selected from the group consisting of glass fiber filters, glass surfaces and metal surfaces.
7. The method of claim 1 , wherein the solid support is selected from the group consisting of steel, gold, sliver, aluminum, copper and silicon.
8. The method of claim 1 , wherein the solid support is silicon.
9. The method of claim 1 , wherein the solid support is a silicon wafer.
10. The method of claim 1 , wherein the oligonucleotides on the solid support are linked to the support via a linker or a bond cleavable under the conditions of mass spectrometric analysis.
11. The method of claim 10 , wherein the oligonucleotides are linked to the support via a linker or bond that is photocleavable.
12. The method of claim 1 , wherein the oligonucleotides comprise oligoribonucleotides, oligodeoxyribonucleotides, nucleotide analogs, or protein nucleic acid (PNA).
13. The method of claim 12 , wherein the nucleotide analogs comprise a thio-modified phosphodiester or phosphotriester backbone.
14. The method of claim 1 , wherein the oligonucleotides are linked to the support by an irreversible bond.
15. The method of claim 1 , wherein the oligonucleotides are linked to the support by a disulfide bond, a biotin/streptavidin linkage, a heterobifunctional derivative of a trityl ether group, a levulinyl group, an arginine-arginine bond, a lysine-lysine bond, a pyrophosphate bond or a charge transfer complex.