We claim:
1. A method for reverse transcribing an RNA, that comprises:
(a) providing a reverse transcription reaction mixture comprising said RNA, a primer, a divalent cation, and a mutant thermoactive DNA polymerase, wherein said mutant DNA polymerase is characterized in that
i) in its native form said DNA polymerase comprises an amino acid sequence that is SEQ ID NO:1;
ii) the amino acid at position 4 of said amino acid sequence is mutated in comparison to said native sequence to an amino acid other than E, A, G, or P; and
(b) treating said reaction mixture at a temperature sufficient for said mutant DNA polymerase to initiate synthesis of an extension product of said primer to provide a cDNA molecule complementary to said RNA.
2. The method of claim 1, wherein said amino acid sequence is SEQ ID NO:2.
3. The method of claim 1, wherein said amino acid sequence is SEQ ID NO:3.
4. The method of claim 1, wherein said amino acid sequence is SEQ ID NO:4.
5. The method of claim 1, wherein said amino acid sequence is SEQ ID NO:5.
6. The method of claim 1, wherein said amino acid sequence is SEQ ID NO:6.
7. The method of claim 1, wherein said amino acid sequence is SEQ ID NO:7.
8. A method of claim 1, wherein said mutant DNA polymerase is thermostable.
9. The method of claim 1, wherein said DNA polymerase is a mutant form of a Thermus species DNA polymerase.
10. The method of claim 1, wherein said DNA polymerase is a mutant form of Thermus thermophilus DNA polymerase or Thermus aquaticus DNA polymerase.
11. The method of claim 1, wherein said temperature of said reaction mixture in step (b) is between 40 C. and 80 C.
12. The method of claim 1, wherein said amino acid at position 4 of said amino acid sequence is mutated in comparison to said native sequence to an amino acid other than E, A, G, P, Q, or D.
13. A method for reverse transcribing an RNA, that comprises:
(a) providing a reverse transcription reaction mixture comprising said RNA, a primer, Mg2, and a mutant thermoactive DNA polymerase, wherein said mutant DNA polymerase is characterized in that
i) in its native form said DNA polymerase comprises an amino acid sequence that is SEQ ID NO:1;
ii) the amino acid at position 4 of said amino acid sequence is mutated in comparison to said native sequence to an amino acid other than E, A, G, or P; and
(b) treating said reaction mixture at a temperature sufficient for said mutant DNA polymerase to initiate synthesis of an extension product of said primer to provide a cDNA molecule complementary to said RNA.
14. The method of claim 13, wherein said amino acid sequence is SEQ ID NO:2.
15. The method of claim 13, wherein said amino acid sequence is SEQ ID NO:3.
16. The method of claim 13, wherein said amino acid sequence is SEQ ID NO:4.
17. The method of claim 13, wherein said amino acid sequence is SEQ ID NO:5.
18. The method of claim 13, wherein said amino acid sequence is SEQ ID NO:6.
19. The method of claim 13, wherein said amino acid sequence is SEQ ID NO:7.
20. The method of claim 13, wherein said mutant DNA polymerase is thermostable.
21. The method of claim 13, wherein said DNA polymerase is a mutant form of a Thermus species DNA polymerase.
22. The method of claim 13, wherein said DNA polymerase is a mutant form of Thermus thermophilus DNA polymerase or Thermus aquaticus DNA polymerase.
23. The method of claim 13, wherein said temperature of said reaction mixture in step (b) is between 40 C. and 80 C.
24. The method of claim 13, wherein said amino acid at position 4 of said amino acid sequence is mutated in comparison to said native sequence to an amino acid other than E, A, G, P, Q, or D.
25. A method for amplifying an RNA, that comprise:
(a) reverse transcribing said RNA according to a method of claim 1 to provide a cDNA;
(b) amplifying said cDNA.
26. A method of claim 25, wherein in step (b) said amplifying is carried out using a polymerase chain reaction.
27. A method for amplifying an RNA, that comprise:
(a) reverse transcribing said RNA according to a method of claim 13 to provide a cDNA;
(b) amplifying said cDNA.
28. A method of claim 27, wherein in step (b) said amplifying is carried out using a polymerase chain reaction.
29. A method for amplifying an RNA using a single-enzyme reverse transcriptionamplification reaction, that comprises:
(a) providing an amplification reaction mixture comprising said RNA, a pair of primers, a divalent cation, and a mutant thermostable DNA polymerase, wherein said mutant DNA polymerase is characterized in that
i) in its native form said DNA polymerase comprises an amino acid sequence that is SEQ ID NO:1;
ii) the amino acid at position 4 of said amino acid sequence is mutated in comparison to said native sequence to an amino acid other than E, A, G, or P; and
(b) treating said reaction mixture at a temperature sufficient for said mutant DNA polymerase to initiate synthesis of an extension product of said primer to provide a cDNA molecule complementary to said RNA;
(c) treating said reaction mixture at an appropriate temperature for said mutant DNA polymerase to initiate synthesis of an extension product of said second primer to provide a double-stranded cDNA molecule; and
(d) amplifying said double-stranded cDNA molecule of step (c) by a polymerase chain reaction.
30. The method of claim 29, wherein said amino acid sequence is SEQ ID NO:2.
31. The method of claim 29, wherein said amino acid sequence is SEQ ID NO:3.
32. The method of claim 29, wherein said amino acid sequence is SEQ ID NO:4.
33. The method of claim 29, wherein said amino acid sequence is SEQ ID NO:5.
34. The method of claim 29, wherein said amino acid sequence is SEQ ID NO:6.
35. The method of claim 29, wherein said amino acid sequence is SEQ ID NO:7.
36. The method of claim 29, wherein said mutant DNA polymerase is thermostable.
37. The method of claim 29, wherein said DNA polymerase is a mutant form of a Thermus species DNA polymerase.
38. The method of claim 29, wherein said DNA polymerase is a mutant form of Thermus thermophilus DNA polymerase or Thermus aquaticus DNA polymerase.
39. The method of claim 29, wherein said temperature of said reaction mixture in step (b) is between 40 C. and 80 C.
40. The method of claim 29, wherein said amino acid at position 4 of said amino acid sequence is mutated in comparison to said native sequence to an amino acid other than E, A, G, P, Q, or D.
41. A method for amplifying an RNA using a single-enzyme reverse transcriptionamplification reaction, that comprises:
(a) providing an amplification reaction mixture comprising said RNA, a pair of primers, Mg2, and a mutant thermostable DNA polymerase, wherein said mutant DNA polymerase is characterized in that
i) in its native form said DNA polymerase comprises an amino acid sequence that is SEQ ID NO:1;
ii) the amino acid at position 4 of said amino acid sequence is mutated in comparison to said native sequence to an amino acid other than E, A, G, or P; and
(b) treating said reaction mixture at a temperature sufficient for said mutant DNA polymerase to initiate synthesis of an extension product of said primer to provide a cDNA molecule complementary to said RNA;
(c) treating said reaction mixture at an appropriate temperature for said mutant DNA polymerase to initiate synthesis of an extension product of said second primer to provide a double-stranded cDNA molecule; and
(d) amplifying said double-stranded cDNA molecule of step (c) by a polymerase chain reaction.
42. The method of claim 41, wherein said amino acid sequence is SEQ ID NO:2.
43. The method of claim 41, wherein said amino acid sequence is SEQ ID NO:3.
44. The method of claim 41, wherein said amino acid sequence is SEQ ID NO:4.
45. The method of claim 41, wherein said amino acid sequence is SEQ ID NO:5.
46. The method of claim 41, wherein said amino acid sequence is SEQ ID NO:6.
47. The method of claim 41, wherein said amino acid sequence is SEQ ID NO:7.
48. The method of claim 41, wherein said mutant DNA polymerase is thermostable.
49. The method of claim 41, wherein said DNA polymerase is a mutant form of a Thermus species DNA polymerase.
50. The method of claim 41, wherein said DNA polymerase is a mutant form of Thermus thermophilus DNA polymerase or Thermus aquaticus DNA polymerase.
51. The method of claim 41, wherein said temperature of said reaction mixture in step (b) is between 40 C. and 80 C.
52. The method of claim 41, wherein said amino acid at position 4 of said amino acid sequence is mutated in comparison to said native sequence to an amino acid other than E, A, G, P, Q, or D.
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 apparatus comprising:
a storage device for storing first and second data tables, each table having one or more fields of which at least some are populated with data entries, both data tables having one or more mutually similar fields and at least one of the data tables having one or more dissimilar fields that are dissimilar to fields of the other data table; and
a processing device in communication with said storage device, said processing device being configured to modify the first and second data tables so as to synchronize the similar fields of the first and second data tables.
2. An apparatus according to claim 1, further comprising a communications device configured to receive from at least one separate device, and to communicate to said storage device, the first and second data tables.
3. An apparatus according to claim 2, wherein the communications device is further configured to communicate at least one of the first and second data tables, subsequent to modification, to at least one separate device.
4. An apparatus according to claim 2, wherein said communications device is further configured to receive from the at least one separate device, and communicate to said storage device, at least one updated data table that replaces at least a respective one of the first and second data tables previously stored in said storage device.
5. An apparatus according to claim 2, wherein said communications device is configured to communicate with at least one separate device that is selected from the group consisting of a mobile phone and a mobile data device.
6. An apparatus according to claim 1, wherein said processing device is further configured to determine a priority between respective differing data entries from similar fields of the first and second data tables and to modify both of the first and second data tables so as to synchronize the similar fields of the first and second data tables by replacing each differing data entry of lower priority with the corresponding differing data entry of higher priority.
7. An apparatus according to claim 6, wherein said storage device includes an intermediate data table having intermediate data table fields corresponding to the mutually similar fields of the first and second data tables, at least some of the intermediate data table fields being populated with previously stored intermediate data table entries, and wherein said processing device is configured to determine a priority and generate a dissimilarity indicator by using data entries of the first and second data tables to respectively replace corresponding inconsistent intermediate data table entries and subsequently using intermediate data table entries to respectively replace corresponding inconsistent first and second data table entries.
8. An apparatus according to claim 6, wherein said storage device includes an intermediate data table having intermediate data table fields corresponding to the mutually similar fields of the first and second data tables, at least some of the intermediate data table fields being populated with previously stored intermediate data table entries, and wherein said storage device includes first and second normalized data tables with respective first and second normalized data table fields corresponding to the mutually similar fields of the first and second data tables, and said processing device is further configured to at least partially populate the first normalized data table with corresponding entries from the first data table and the second normalized data table with corresponding entries in the second data table, and wherein said processing unit is configured to determine a priority and generate a dissimilarity indicator by using data entries of the first and second normalized data tables to respectively replace corresponding inconsistent intermediate data table entries and subsequently using intermediate data table entries to respectively replace corresponding inconsistent first and second normalized data table entries.
9. An apparatus according to claim 8, wherein the entries in the first and second normalized data tables and in the intermediate data table are in a common data format, and wherein the processing device is further configured to use the entries in the first and second normalized data tables in common data format to generate data in alternative formats compatible with an external device.
10-20. (canceled)