US 9,951,378 C1 (12,974th)
Compositions, methods and kits for real time polymerase chain reaction (PCR)
Carole Bornarth, Fremont, CA (US); Michael Lau, Sunnyvale, CA (US); and Junko Stevens, Menlo Park, CA (US)
Filed by LIFE TECHNOLOGIES CORPORATION, Carlsbad, CA (US)
Assigned to LIFE TECHNOLOGIES CORPORATION, Carlsbad, CA (US)
Reexamination Request No. 90/019,445, Apr. 11, 2024.
Reexamination Certificate for Patent 9,951,378, issued Apr. 24, 2018, Appl. No. 13/918,768, Jun. 14, 2013.
Claims priority of provisional application 61/659,587, filed on Jun. 14, 2012.
Ex Parte Reexamination Certificate issued on Jul. 9, 2025.
Int. Cl. C12Q 1/6848 (2018.01); C07K 16/00 (2006.01); C12N 9/00 (2006.01); C12Q 1/686 (2018.01)
CPC C12Q 1/6848 (2013.01) [C07K 16/00 (2013.01); C12N 9/00 (2013.01); C12Q 1/686 (2013.01)]
AS A RESULT OF REEXAMINATION, IT HAS BEEN DETERMINED THAT:
Claims 5 and 9 are cancelled.
Claims 1 and 6 are determined to be patentable as amended.
Claims 2-4, 7, 8 and 13-15, dependent on an amended claim, are determined to be patentable.
New claims 23-29 and 42-47 are added and determined to be patentable.
Claims 10-12 and 16-22 were not reexamined.
1. A composition comprising:
a) a nucleic acid polymerase;
b) at least one nucleic acid binding dye;
c) a first hot start mechanism comprising antibodies or combinations of antibodies specific to the polymerase; and
d) a second hot start mechanism [ comprising at least one selected from the group consisting of oligonucleotides that block activity of the polymerase at lower temperatures, reversible chemical modifications of the nucleic acid polymerase that dissociate at elevated temperatures, and amino acid modifications of the nucleic acid polymerase that provide reduced activity at lower temperatures] ,
wherein the first hot start mechanism and the second hot start mechanism are different and which, together, are capable of substantially inhibiting [ the ] activity of the polymerase at a first temperature, but not at a second temperature.
6. A composition comprising:
a) a first thermostable nucleic acid polymerase;
b) at least one nucleic acid binding dye; and
c) at least two different hot start mechanisms which, together, are capable of inhibiting the first thermostable nucleic acid polymerase;
wherein one of the at least two different hot start mechanisms comprises antibody-mediated inhibition of the thermostable nucleic acid polymerase ; [ ,
wherein another of the least two different hot start mechanisms comprises at least one selected from the group consisting of oligonucleotides that block the activity of the first thermostable nucleic acid polymerase at lower temperatures, reversible chemical modifications of the first thermostable nucleic acid polymerase that dissociate at elevated temperatures, and amino acid modifications of the first thermostable nucleic acid polymerase that provide reduced activity at lower temperatures,]
wherein the at least two hot start mechanisms substantially inhibit the polymerase activity of the thermostable nucleic acid polymerase at a temperature less than about 40° C.; and
wherein the at least two hot start mechanisms do not substantially inhibit the polymerase activity of the first thermostable nucleic acid polymerase at a temperature greater than about 40° C.
[ 23. The composition of claim 1, wherein the first hot start mechanism and the second hot start mechanism together are capable of substantially inhibiting activity of the polymerase for at least 24 hours at a first temperature, but not at a second temperature, wherein the first temperature is about 20° C. to about 40° C.]
[ 24. The composition of claim 1, wherein the second hot start mechanism comprises oligonucleotides that block polymerase activity at lower temperatures.]
[ 25. The composition of claim 24, wherein the second hot start mechanism comprises oligonucleotides that block polymerase activity at about 20° C.]
[ 26. The composition of claim 1, wherein the second hot start mechanism comprises reversible chemical modifications of the nucleic acid polymerase that dissociate at elevated temperatures.]
[ 27. The composition of claim 26, wherein the second hot start mechanism comprises reversible chemical modifications of the nucleic acid polymerase that dissociate at greater than 50° C.]
[ 28. The composition of claim 1, wherein the second hot start mechanism comprises amino acid modifications of the nucleic acid polymerase that provide reduced activity at lower temperatures.]
[ 29. The composition of claim 28, wherein the second hot start mechanism comprises amino acid modifications of the nucleic acid polymerase that provide reduced activity at about 20° C.]
[ 30. The composition of claim 6, wherein the second hot start mechanism comprises oligonucleotides that block polymerase activity at lower temperatures.]
[ 31. The composition of claim 30, wherein the second hot start mechanism comprises oligonucleotides that block polymerase activity at about 20° C.]
[ 32. The composition of claim 6, wherein the second hot start mechanism comprises reversible chemical modifications of the nucleic acid polymerase that dissociate at elevated temperatures.]
[ 33. The composition of claim 32, wherein the second hot start mechanism comprises reversible chemical modifications of the nucleic acid polymerase that dissociate at greater than 50° C.]
[ 34. The composition of claim 6, wherein the second hot start mechanism comprises amino acid modifications of the nucleic acid polymerase that provide reduced activity at lower temperatures.]
[ 35. The composition of claim 34, wherein the second hot start mechanism comprises amino acid modifications of the nucleic acid polymerase that provide reduced activity at about 20° C.]