MODIFIED THERMOCCOCUS POLYMERASES

1 . A polymerase comprising an amino acid sequence that is at least 80% identical to a continuous 500 amino acid sequence within SEQ ID NO: 1, wherein the polymerase comprises:

an alanine or serine at amino acid position 411 or an amino acid functionally equivalent to amino acid position 411; and

an amino acid substitution, wherein said amino acid substitution comprises: P36A, P36H, V93Q, F152E, G155T, D215A, D317A, or K480A.

2 . The polymerase of claim 1 , further comprising an alanine or glycine at amino acid position 93 or an amino acid functionally equivalent to amino acid position 93.

3 . The polymerase of claim 1 , further comprising at least one of the following amino acid substitutions:

an alanine at amino acid position 129 or an amino acid position functionally equivalent to amino acid position 129;

an alanine at amino acid position 141 or an amino acid position functionally equivalent to amino acid position 141;

an alanine at amino acid position 143 or an amino acid position functionally equivalent to amino acid position 143;

an alanine at amino acid position 144 or an amino acid position functionally equivalent to amino acid position 144;

a glutamic acid or threonine at amino acid position 153 or an amino acid position functionally equivalent to amino acid position 153;

an aspartic acid at amino acid position 157 or an amino acid position functionally equivalent to amino acid position 157;

an isoleucine at amino acid position 303 or an amino acid position functionally equivalent to amino acid position 303;

a methionine at amino acid position 307 or an amino acid position functionally equivalent to amino acid position 307;

a phenylalanine at amino acid position 381 or an amino acid position functionally equivalent to amino acid position 381;

a tryptophan at amino acid position 479 or an amino acid position functionally equivalent to amino acid position 479;

an alanine at amino acid position 481 or an amino acid position functionally equivalent to amino acid position 481;

a valine at amino acid position 488 or an amino acid position functionally equivalent to amino acid position 488;

a serine at amino acid position 517 or an amino acid position functionally equivalent to amino acid position 517;

an isoleucine at amino acid position 593 or an amino acid position functionally equivalent to amino acid position 593;

an alanine at amino acid position 605 or an amino acid position functionally equivalent to amino acid position 605; or

a leucine at amino acid position 642 or an amino acid position functionally equivalent to amino acid position 642.

4 . The polymerase of claim 1 , wherein the polymerase comprises an alanine at amino acid position 411 or an amino acid functionally equivalent to amino acid position 411.

5 . The polymerase of claim 1 , wherein the polymerase comprises a serine at amino acid position 411 or an amino acid functionally equivalent to amino acid position 411.

6 . The polymerase of claim 1 , further comprising a glycine at amino acid position 412 or an amino acid functionally equivalent to amino acid position 412.

7 . The polymerase of claim 1 , further comprising an alanine at amino acid position 412 or an amino acid functionally equivalent to amino acid position 412.

8 . The polymerase of claim 1 , further comprising a proline at amino acid position 413 or an amino acid functionally equivalent to amino acid position 413.

9 . The polymerase of claim 1 , further comprising a valine at amino acid position 413 or an amino acid functionally equivalent to amino acid position 413.

10 . The polymerase of claim 1 , further comprising an isoleucine at amino acid position 413 or an amino acid functionally equivalent to amino acid position 413.

11 . The polymerase of claim 1 , which exhibits an increased rate of incorporation of modified nucleotides, relative to a control.

12 . The polymerase of claim 1 , which is capable of incorporating modified nucleotides at reaction temperatures across the range of 40° C. to 80° C.

13 . The polymerase of claim 1 , which exhibits an exonuclease activity that is reduced by at least 80% relative to the exonuclease activity of a polymerase of SEQ ID NO: 1.

14 . The polymerase of claim 1 , comprising the amino acid substitutions:

L411A, Y412G, F152E;

L411A, Y412G, G155T;

L411A, Y412G, P36H;

L411A, Y412G, P36A;

L411A, Y412G, V93G;

L411A, Y412G, V93A;

L411A, Y412G, V93Q;

D141A, E143A, L411A, Y412G, F152E;

D141A, E143A, L411A, Y412G, G155T;

D141A, E143A, L411A, Y412G, P36H;

D141A, E143A, L411A, Y412G, P36A;

D141A, E143A, L411A, Y412G, V93G;

D141A, E143A, L411A, Y412G, V93A;

D141A, E143A, L411A, Y412G, V93Q;

L411A, Y412A, P413I, F152E;

L411A, Y412A, P413I, G155T;

L411A, Y412A, P413I, P36H;

L411A, Y412A, P413I, P36A;

L411A, Y412A, P413I, V93G;

L411A, Y412A, P413I, V93A;

L411A, Y412A, P413I, V93Q;

M129A, D141A, E143A, L411A, Y412A, P413I, F152E;

M129A, D141A, E143A, L411A, Y412A, P413I, G155T;

M129A, D141A, E143A, L411A, Y412A, P413I, P36H;

M129A, D141A, E143A, L411A, Y412A, P413I, P36A;

M129A, D141A, E143A, L411A, Y412A, P413I, V93G;

M129A, D141A, E143A, L411A, Y412A, P413I, V93A; or

M129A, D141A, E143A, L411A, Y412A, P413I, V93Q.

15 . A method of extending a primer, said method comprising contacting a primer hybridized to a template polynucleotide with a nucleotide and extending said primer with a polymerase by incorporating the modified nucleotide to form an extended primer, wherein the polymerase is the polymerase of claim 1 .

16 . A method of incorporating a modified nucleotide into a nucleic acid sequence comprising allowing the following components to interact: (i) a DNA template, (ii) a nucleotide solution, and (iii) the polymerase of claim 1 .

17 . A method of sequencing a nucleic acid sequence comprising:

(i) hybridizing a nucleic acid template with a primer to form a primer-template hybridization complex;

(ii) contacting the primer-template hybridization complex with a DNA polymerase and nucleotides, wherein the DNA polymerase is a polymerase of claim 1 and the nucleotides comprise a modified nucleotide, wherein the modified nucleotide comprises a detectable label;

(iii) subjecting the primer-template hybridization complex to conditions which enable the polymerase to incorporate the modified nucleotide into the primer-template hybridization complex to form a modified primer-template hybridization complex; and

(iv) detecting the detectable label; thereby sequencing the nucleic acid sequence.