NANOPORE SEQUENCING USING CHARGE BLOCKADE LABELS
First Claim
1. A composition comprising:
- a plurality of polymerase enzymes, each complexed with a template nucleic acid, each polymerase enzyme attached to a nanopore or attached proximal to a nanopore, and nucleic acid sequencing reagents including at least one nucleotide analog having the structure NS-PP-L-B wherein NS comprise a nucleoside moiety, PP comprises a polyphosphate chain with at least two phosphates, L comprises a linker, and B comprises a blockade label.
0 Assignments
0 Petitions
Accused Products
Abstract
The invention relates to devices and methods for nanopore sequencing. The invention includes compositions and methods of nucleic acid sequencing using a single polymerase enzyme complex comprising a polymerase enzyme and a template nucleic acid attached proximal to a nanopore, and nucleotide analogs in solution comprising charge blockade label that are attached to the polyphosphate portion of the nucleotide analog such that the charge blockade labels are cleaved when the nucleotide analog is incorporated into a growing nucleic acid and the charge blockade label is detected by the nanopore to determine the presence and identity of the incorporated nucleotide and thereby determine the sequence of a template nucleic acid.
-
Citations
20 Claims
-
1. A composition comprising:
- a plurality of polymerase enzymes, each complexed with a template nucleic acid, each polymerase enzyme attached to a nanopore or attached proximal to a nanopore, and nucleic acid sequencing reagents including at least one nucleotide analog having the structure NS-PP-L-B wherein NS comprise a nucleoside moiety, PP comprises a polyphosphate chain with at least two phosphates, L comprises a linker, and B comprises a blockade label.
- View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14)
-
15. A composition comprising:
- a plurality of DNA polymerase enzymes, each complexed with a primed template DNA, each DNA polymerase enzyme attached to a protein nanopore, and nucleic acid sequencing reagents including four different nucleotide analogs corresponding to A, G, C, T, or A, G, C, U, each nucleotide analog having the structure NS-PP-L-B wherein NS comprise a nucleoside moiety, PP comprises a polyphosphate chain with at least two phosphates, L comprises a linker, and B comprises a blockade label.
- View Dependent Claims (16, 17, 18, 19, 20)
Specification