Modified Proteins and Methods of Making and Using Same
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Abstract
Methods, compositions, systems, apparatuses and kits comprising modified proteins, particularly modified nucleic acid-binding proteins with altered buffering properties are provided. For example, in some embodiments, methods of forming modified proteins including one or more amino acid modifications to achieve desired pKa values are described. Furthermore, the invention provides methods for using such modified proteins in ion-producing reactions, such as ion-based nucleic acid sequencing reactions.
10 Citations
29 Claims
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1-15. -15. (canceled)
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16. An apparatus for nucleotide polymerization, comprising:
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(a) a field effect transistor (FET); and (b) a modified polymerase including one or more amino acid substitutions that reduce the buffering capacity of the modified polymerase relative to the corresponding unmodified polymerase, wherein the polymerase is configured to catalyze nucleotide polymerization; and wherein the FET is configured to detect the release of a chemical byproduct of nucleotide incorporation catalyzed by the modified polymerase. - View Dependent Claims (17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29)
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Specification