1. An enzyme mixture comprising a first enzyme and a second enzyme, wherein said first enzyme comprises 5\u2032-3\u2032 polymerization activity of a DNA polymerase or reverse transcriptase, and said second enzyme is a mutant of a wild type DNA polymerase, said wild type DNA polymerase comprising the partitioning domain sequence YXGG (SEQ ID NO:6), the polymerase domain sequence DXXSLYP (SEQ ID NO:1) or DFRALYP (SEQ ID NO:13), the polymerase domain sequence YXDTDS (SEQ ID NO:4), YIDTDG (SEQ ID NO:15), YADTDG (SEQ ID NO:16), or YSDTDG (SEQ ID NO:17), and the polymerase domain sequence KXY, wherein said second enzyme comprises an amino acid substitution at an amino acid position corresponding to G387 of SEQ ID NO:19, and wherein said second enzyme comprises 3\u2032-5\u2032 exonuclease activity and reduced 5\u2032-3\u2032 DNA polymerization activity as compared to the wild type DNA polymerase.
2. The enzyme mixture of claim 1, wherein said first enzyme is a DNA polymerase or a reverse transcriptase.
3. The enzyme mixture of claim 2, wherein said first enzyme is selected from the group consisting of: Taq DNA polymerase, Tth DNA polymerase, UlTma DNA polymerase, Tli DNA polymerase, Pfu DNA polymerase, KOD DNA polymerase, JDF-3 DNA polymerase, PGB-D DNA polymerase and DP1DP2 DNA polymerase.
4. The enzyme mixture of claim 1, wherein said first enzyme is a Pfu DNA polymerase.
5. The enzyme mixture of claim 1, further comprising a PCR enhancing factor andor an additive.
6. The enzyme mixture of claim 1, wherein said enzyme mixture has a ratio of polymerization activityexonuclease activity of (2.5-5U)(0.02-5U).
7. The enzyme mixture of claim 6, wherein said enzyme mixture has a ratio of polymerization activityexonuclease activity of (2.5U)(0.04-0.08U).
8. An enzyme mixture comprising three or more enzymes, wherein at least one enzyme in said enzyme mixture is a mutant of a wild type DNA polymerase, said wild type DNA polymerase comprising the partitioning domain sequence YXGG (SEQ ID NO:6), the polymerase domain sequence DXXSLYP (SEQ ID NO:1) or DFRALYP (SEQ ID NO:13), the polymerase domain sequence YXDTDS (SEQ ID NO:4), YIDTDG (SEQ ID NO:15), YADTDG (SEQ ID NO:16), or YSDTDG (SEQ ID NO:17), and the polymerase domain sequence KXY, wherein said mutant DNA polymerase comprises 3\u2032-5\u2032 exonuclease activity and a reduced DNA polymerization activity as compared to the wild type DNA polymerase, and wherein said mutant DNA polymerase comprises an amino acid substitution at an amino acid position corresponding to G387 of SEQ ID NO:19.
9. The enzyme mixture of claim 8, wherein at least two enzymes in said mixture are mixed as an enzyme blend before being added to said enzyme mixture.
10. The enzyme mixture of claim 9, wherein said enzyme blend comprises a Pfu DNA polymerase and Taq DNA polymerase.
11. The enzyme mixture of claim 10, wherein said enzyme blend further comprises a PCR enhancing factor.
12. A kit comprising a first enzyme, a second enzyme, and packaging material therefor, wherein said first enzyme is a DNA polymerase or reverse transcriptase comprising 5\u2032-3\u2032 polymerization activity of a DNA polymerase or reverse transcriptase, said second enzyme is a mutant of a wild type DNA polymerase, said wild type DNA polymerase comprising the partitioning domain sequence YXGG (SEQ ID NO:6), the polymerase domain sequence DXXSLYP (SEQ ID NO:1) or DFRALYP (SEQ ID NO:13), the polymerase domain sequence YXDTDS (SEQ ID NO:4), YIDTDG (SEQ ID NO:15), YADTDG (SEQ ID NO:16), or YSDTDG (SEQ ID NO:17), and the polymerase domain sequence KXY, wherein said second enzyme comprises an amino acid substitution at an amino acid position corresponding to G387 of SEQ ID NO:19 and wherein said second enzyme comprises 3\u2032-5\u2032 exonuclease activity and a reduced DNA polymerization activity as compared to the wild type DNA polymerase.
13. The kit of claim 12, wherein said first enzyme is selected from the group consisting of: Taq DNA polymerase, Tth DNA polymerase, UlTma DNA polymerase, Tli DNA polymerase, Pfu DNA polymerase, KOD DNA polymerase, JDF-3 DNA polymerase, PGB-D DNA polymerase and DP1DP2 DNA polymerase.
14. The kit of claim 12, wherein said first enzyme is a Pfu DNA polymerase.
15. The kit of claim 12 or 14, further comprising one or more components selected from the group consisting of: a deoxynucleotide, a reaction buffer, a PCR enhancing factor andor an additive, a control DNA template and a control primer.
16. The kit of claim 12, wherein said enzyme mixture has a ratio of polymerization activityexonuclease activity of (2.5-5U)(0.02-5U).
17. The kit of claim 16, wherein said enzyme mixture has a ratio of polymerization activityexonuclease activity of (2.5U)(0.04-0.08U).
18. The enzyme mixture of claim 1, wherein said first enzyme is Taq DNA polymerase or Pfu DNA polymerase.
19. The kit of claim 12, wherein said first enzyme is Taq DNA polymerase or Pfu DNA polymerase.
20. The enzyme mixture of claim 1 or 4, wherein said second enzyme is a mutant DNA polymerase from the species Pyrococcus furiosis.
21. The enzyme mixture of claim 8, wherein said at least one enzyme is a mutant DNA polymerase from the species Pyrococcus furiosis.
22. The kit of claim 12 or 14, wherein said second enzyme is a mutant DNA polymerase from the species Pyrococcus furiosis.
23. The enzyme mixture of claim 1, wherein said wild type DNA polymerase is a Family B DNA polymerase.
24. The enzyme mixture of claim 1, wherein said wild type DNA polymerase is selected from the group consisting of Thermococcus litoralis DNA polymerase (Vent DNA polymerase) having the amino acid sequence of SEQ ID NO:49, Pyrococcus GB-D (Deep Vent) DNA polymerase having the amino acid sequence of SEQ ID NO:59, Thermococcus gorgonarius DNA polymerase having the amino acid sequence of SEQ ID NO:29, Pyrococcus furiousus DNA polymerase having the amino acid sequence of SEQ ID NO:19, and Pyrococcus kodakaraensis DNA polymerase having the amino acid sequence of SEQ ID NO:39.
25. The enzyme mixture of claim 24, wherein said second enzyme comprises an amino acid substitution at G387 in the amino acid sequence of SEQ ID NO:19.
26. The enzyme mixture of claim 24, wherein said second enzyme comprises an amino acid substitution at G386 in the amino acid sequence of SEQ ID NO:29.
27. The enzyme mixture of claim 24, wherein said second enzyme comprises an amino acid substitution at G386 in the amino acid sequence of SEQ ID NO:39.
28. The enzyme mixture of claim 24, wherein said second enzyme comprises an amino acid substitution at G389 in the amino acid sequence of SEQ ID NO:49.
29. The enzyme mixture of claim 24, wherein said second enzyme comprises an amino acid substitution at G387 in the amino acid sequence of SEQ ID NO:59.
30. The enzyme mixture of claim 23, wherein said second enzyme comprises the amino acid sequence of SEQ ID NO:19 except for an amino acid substitution at G387.
31. The kit of claim 12, wherein said wild type DNA polymerase is a Family B DNA polymerase.
32. The kit of claim 12, wherein said wild type DNA polymerase is selected from the group consisting of Thermococcus litoralis DNA polymerase (Vent DNA polymerase) having the amino acid sequence of SEQ ID NO:49, Pyrococcus GB-D (Deep Vent) DNA polymerase having the amino acid sequence of SEQ ID NO:59, Thermococcus gorgonarius DNA polymerase having the amino acid sequence of SEQ ID NO:29, Pyrococcus furiousus DNA polymerase having the amino acid sequence of SEQ ID NO:19, and Pyrococcus kodakaraensis DNA polymerase having the amino acid sequence of SEQ ID NO:39.
33. The kit of claim 32, wherein said second enzyme comprises an amino acid substitution at G387 in the amino acid sequence of SEQ ID NO:19.
34. The kit of claim 32, wherein said second enzyme comprises an amino acid substitution at G386 in the amino acid sequence of SEQ ID NO:29.
35. The kit of claim 32, wherein said second enzyme comprises an amino acid substitution at G386 in the amino acid sequence of SEQ ID NO:39.
36. The kit of claim 32, wherein said second enzyme comprises an amino acid substitution at G389 in the amino acid sequence of SEQ ID NO:49.
37. The kit of claim 32, wherein said second enzyme comprises an amino acid substitution at G387 in the amino acid sequence of SEQ ID NO:59.
38. The kit of claim 32, wherein said second enzyme comprises the amino acid sequence of SEQ ID NO:19 except for an amino acid substitution at G387.
39. The enzyme mixture of claim 25, wherein the amino acid substitution at G3 87 is a glycine to proline substitution.
40. The enzyme mixture of claim 30, wherein the amino acid substitution at G3 87 is a glycine to proline substitution.
41. The kit of claim 33, wherein the amino acid substitution at G387 is a glycine to proline substitution.
42. The kit of claim 38, wherein the amino acid substitution at G387 is a glycine to proline substitution.
43. The enzyme mixture of claim 39, wherein the first enzyme is Taq DNA polymerase or Pfu DNA polymerase.
44. The enzyme mixture of claim 40, wherein the first enzyme is Taq DNA polymerase or Pfu DNA polymerase.
45. The kit of claim 41, wherein the first enzyme is Taq DNA polymerase or Pfu DNA polymerase.
46. The kit of claim 42, wherein the first enzyme is Taq DNA polymerase or Pfu DNA polymerase.
47. The enzyme mixture of claim 43, wherein the first enzyme is Taq DNA polymerase.
48. The enzyme mixture of claim 44, wherein the first enzyme is Taq DNA polymerase.
49. The kit of claim 45, wherein the first enzyme is Taq DNA polymerase.
50. The kit of claim 46, wherein the first enzyme is Taq DNA polymerase.
The claims below are in addition to those above.
All refrences to claim(s) which appear below refer to the numbering after this setence.
1. An adhesive composition whose main component is a polymer prepared by copolymerizing a monomer composition containing styrene, a (meth)acrylic acid ester having a cyclic structure, and an alkyl(meth)acrylate having a chain structure,
the monomer composition further containing a carboxylic acid having an ethylenic double bond, a bifunctional monomer having two functional groups, and a styrene macromonomer, and
the polymer having a styrene block segment.
2. The adhesive composition as set forth in claim 1, wherein:
the amount of carboxylic acid is 1 to 10 parts by mass, based on 100 parts by mass of the styrene, the (meth)acrylic acid ester, and the alkyl(meth)acrylate.
3. The adhesive composition as set forth in claim 1, wherein:
the carboxylic acid is represented by Formula (1):
R1COOH)m\u2003\u2003(1)
wherein R1 is a C2 to C20 organic group having a (meth)acryloyl group or a vinyl group; and m represents an integer of 1 to 3.
4. The adhesive composition as set forth in claim 3, wherein R1 has an oxygen atom.
5. The adhesive composition as set forth in claim 1, wherein:
the carboxylic acid is (meth)acrylic acid or a carboxylic acid represented by Formula (2):
where R2 is a hydrogen atom or a C1 to C4 alkyl group; and R3 is a divalent C1 to C5 alkylene group or a divalent C4 to C20 organic group having a cyclic structure.
6. The adhesive composition as set forth in claim 5, wherein R2 or R3 has an oxygen atom.
7. The adhesive composition as set forth in claim 1, wherein:
the bifunctional monomer has two functional groups selected from the group consisting of a (meth)acryloyl group and a vinyl group.
8. The adhesive composition as set forth in claim 1, wherein:
the bifunctional monomer is at least one monomer selected from compounds represented by Formula (3):
X1\u2014R4\u2014X2\u2003\u2003(3)
wherein R4 is an organic group selected from a divalent C2 to C20 alkyl group and a divalent C6 to C20 organic group having a cyclic structure, and X1 and X2 are independently a (meth)acryloyl group or a vinyl group.
9. The adhesive composition as set forth in claim 8, wherein R4 has an oxygen atom.
10. The adhesive composition as set forth in claim 1, wherein:
the bifunctional monomer is at least one functional monomer selected from the group consisting of dimethylol-tricyclodecane diacrylate, neopentyl glycol diacrylate, 1,9-nonanediol acrylate, naphthalene diacrylate, and compounds represented by Formula (4):
wherein R5 and R6 are independently ethylene oxide or propylene oxide; and n and S independently represent an integer of 0 to 4.
11. The adhesive composition as set forth in claim 1, wherein:
the amount of bifunctional monomer is 0.1 to 0.5 parts by mass, based on 100 parts by mass of the styrene, the (meth)acrylic acid ester, and the alkyl(meth)acrylate.
12. The adhesive composition as set forth in claim 1, wherein:
the styrene macromonomer has a styrene block structure and organic groups positioned on both ends of the styrene block structure, and
at least one of the organic groups has a carbon-carbon double bond.
13. The adhesive composition as set forth in claim 1, wherein:
the styrene macromonomer is represented by Formula (5):
wherein R7 and R8 are each independently selected from the group consisting of C1 to C10 organic groups having at least one carbon-carbon double bond.
14. The adhesive composition as set forth in claim 13, wherein R7 or R8 has an oxygen atom.
15. The adhesive composition as set forth in claim 1, wherein:
a total amount of the styrene and the styrene macromonomer is 30 to 90 parts by mass, based on 100 parts by mass of the monomer composition, and the amount of styrene macromonomer is 5 to 40 parts by mass, based on the total mass of the styrene and the styrene macromonomer.
16. A film adhesive comprising:
a film; and
an adhesive layer on the film, containing an adhesive composition as set forth in claim 1.