Method for nucleic acid sequencing
AssigneeNagayama IP Holdings LLC
InventorKuniaki Nagayama
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Abstract
A method for determining the sequence of at least a portion of a single-stranded nucleic acid molecule by base-specifically labeling the exposed bases of the nucleic acid molecule using heavy element-substituted nucleotide bases which form Watson-Crick type base-pairs with the exposed bases of the nucleic acid molecule and then imaging the labeled single-stranded nucleic acid molecule using electron microscopy, e.g., transmission electron microscopy (TEM), or some other method that permits discrimination of the heavy element substituted nucleotide bases is described. The image is analyzed to determine the base sequence of at least a portion of the nucleic acid molecule.
Claims37
A method for determining the sequence of a plurality of bases in a nucleic acid molecule, comprising:
- (a) providing a support surface bearing a nucleic acid molecule having a single stranded region;
- (b) exposing the nucleic acid molecule to four different base-specific nucleotides in the presence of an organic solvent, wherein each of the four different base-specific nucleotides comprises a different element having an atomic number greater than 25 and each element having an atomic number greater than 25 differs in atomic number from each other element having an atomic number greater than 25 by an atomic number of at least 15;
- (c) allowing the four different base-specific nucleotides to form base-pairs with bases in the single stranded region of the nucleic acid molecule;
- (d) removing at the non-base paired base-specific nucleotides by washing;
- (e) detecting the base paired nucleotides by electron microscopy;
- and
- (f) analyzing the detected base paired nucleotides to determine the sequence of a plurality of bases in the nucleic acid molecule.
Dependencies are inferred from the claim text.