Abstract: The present invention relates to the field of ligases. More specifically it relates to novel and highly efficient ATP-dependent DNA ligases with a unique ligase activity making the ligase particularly useful in a variety of molecular biology techniques. Furthermore, the invention relates to compositions and kits comprising the DNA ligase, methods for its manufacture and use.

Abstract: The invention provides a composition comprising a proteinase or an enzymatically active fragment thereof, said proteinase comprising the amino acid sequence of SEQ ID NO: 1 or comprising an amino acid sequence which is at least about 70% identical to SEQ ID NO: 1, wherein i) the concentration of free calcium in said composition is ?about 80 ?M; or ii) the concentration of monovalent salt in said composition is ?about 20 mM. Under such conditions, the proteinases and enzymatically active fragments thereof are inducibly thermolabile. The invention further provides samples comprising one or more polypeptides and a proteinase or an enzymatically active fragment thereof, said proteinase comprising the amino acid sequence of SEQ ID NO: 1 or comprising an amino acid sequence which is at least about 70% identical to SEQ ID NO: 1, wherein i) the concentration of free calcium in said sample is ?about 80 ?M; or ii) the concentration of monovalent salt in said sample is ?about 20 mM.

Abstract: The invention provides a composition comprising a proteinase or an enzymatically active fragment thereof, said proteinase comprising the amino acid sequence of SEQ ID NO: 1 or comprising an amino acid sequence which is at least about 70% identical to SEQ ID NO: 1, wherein i) the concentration of free calcium in said composition is ?about 80 ?M; or ii) the concentration of monovalent salt in said composition is ?about 20 mM. Under such conditions, the proteinases and enzymatically active fragments thereof are inducibly thermolabile. The invention further provides samples comprising one or more polypeptides and a proteinase or an enzymatically active fragment thereof, said proteinase comprising the amino acid sequence of SEQ ID NO: 1 or comprising an amino acid sequence which is at least about 70% identical to SEQ ID NO: 1, wherein i) the concentration of free calcium in said sample is ?about 80 ?M; or ii) the concentration of monovalent salt in said sample is ?about 20 mM.

Abstract: The invention provides an exonuclease or an enzymatically active fragment thereof, said exonuclease having the amino acid sequence of SEQ ID NO:1 or an amino acid sequence which is at least about 50% identical thereto, wherein said exonuclease or enzymatically active fragment thereof (i) is substantially irreversibly inactivated by heating at a temperature of about 55° C. for 10 minutes in a buffer consisting of 10 mM Tris-HCl, pH 8.5 at 25° C., 50 mM KCl and 5 mM MgCl2; (ii) is substantially specific for single stranded DNA; and (iii) has a 3?-5? exonuclease activity. The invention further provides a method of removing single stranded DNA from a sample, a method of nucleic acid amplification, a method of reverse transcription and a method of nucleic acid sequence analysis in which the exonuclease or enzymatically active fragment thereof is used.

Abstract: The invention provides an exonuclease or an enzymatically active fragment thereof, said exonuclease having the amino acid sequence of SEQ ID NO:1 or an amino acid sequence which is at least about 50% identical thereto, wherein said exonuclease or enzymatically active fragment thereof (i) is substantially irreversibly inactivated by heating at a temperature of about 55° C. for 10 minutes in a buffer consisting of 10 mM Tris-HCl, pH 8.5 at 25° C., 50 mM KCl and 5 mM MgCl2; (ii) is substantially specific for single stranded DNA; and (iii) has a 3?-5? exonuclease activity. The invention further provides a method of removing single stranded DNA from a sample, a method of nucleic acid amplification, a method of reverse transcription and a method of nucleic acid sequence analysis in which the exonuclease or enzymatically active fragment thereof is used.

Abstract: The present invention relates to DNA polymerases. In particular, the present invention relates to DNA polymerases based on a DNA polymerase from a Psychrobacillus sp. The present invention provides an isolated DNA polymerase or an enzymatically active fragment thereof, said DNA polymerase comprising the amino acid sequence of SEQ ID NO:1 or comprising an amino acid sequence which is at least 70% identical to SEQ ID NO:1. The invention also provides nucleic acid molecules comprising a nucleotide sequence that encodes the DNA polymerase. The invention also provides a method of nucleotide polymerisation and a method of amplifying a nucleic acid in which the DNA polymerase or an enzymatically active fragment thereof is used.

Abstract: The invention provides an exonuclease or an enzymatically active fragment thereof, said exonuclease having the amino acid sequence of SEQ ID NO:1 or an amino acid sequence which is at least about 50% identical thereto, wherein said exonuclease or enzymatically active fragment thereof (i) is substantially irreversibly inactivated by heating at a temperature of about 55° C. for 10 minutes in a buffer consisting of 10 mM Tris-HCl, pH 8.5 at 25° C., 50 mM KCl and 5 mM MgCl2; (ii) is substantially specific for single stranded DNA; and (iii) has a 3?-5? exonuclease activity. The invention further provides a method of removing single stranded DNA from a sample, a method of nucleic acid amplification, a method of reverse transcription and a method of nucleic acid sequence analysis in which the exonuclease or enzymatically active fragment thereof is used.

Abstract: The invention provides an exonuclease or an enzymatically active fragment thereof, said exonuclease having the amino acid sequence of SEQ ID NO:1 or an amino acid sequence which is at least about 50% identical thereto, wherein said exonuclease or enzymatically active fragment thereof (i) is substantially irreversibly inactivated by heating at a temperature of about 55° C. for 10 minutes in a buffer consisting of 10 mM Tris-HCl, pH 8.5 at 25° C., 50 mM KCl and 5 mM MgCl2; (ii) is substantially specific for single stranded DNA; and (iii) has a 3?-5? exonuclease activity. The invention further provides a method of removing single stranded DNA from a sample, a method of nucleic acid amplification, a method of reverse transcription and a method of nucleic acid sequence analysis in which the exonuclease or enzymatically active fragment thereof is used.

Abstract: The present invention relates to DNA polymerases. In particular, the present invention relates to DNA polymerases based on a DNA polymerase from a Psychrobacillus sp. The present invention provides an isolated DNA polymerase or an enzymatically active fragment thereof, said DNA polymerase comprising the amino acid sequence of SEQ ID NO:1 or comprising an amino acid sequence which is at least 70% identical to SEQ ID NO:1. The invention also provides nucleic acid molecules comprising a nucleotide sequence that encodes the DNA polymerase. The invention also provides a method of nucleotide polymerisation and a method of amplifying a nucleic acid in which the DNA polymerase or an enzymatically active fragment thereof is used.

Abstract: The invention provides an exonuclease or an enzymatically active fragment thereof, said exonuclease having the amino acid sequence of SEQ ID NO:1 or an amino acid sequence which is at least about 50% identical thereto, wherein said exonuclease or enzymatically active fragment thereof (i) is substantially irreversibly inactivated by heating at a temperature of about 55° C. for 10 minutes in a buffer consisting of 10 mM Tris-HCl, pH 8.5 at 25° C., 50 mM KCl and 5 mM MgCl2; (ii) is substantially specific for single stranded DNA; and (iii) has a 3?-5? exonuclease activity. The invention further provides a method of removing single stranded DNA from a sample, a method of nucleic acid amplification, a method of reverse transcription and a method of nucleic acid sequence analysis in which the exonuclease or enzymatically active fragment thereof is used.

Abstract: The invention provides an exonuclease or an enzymatically active fragment thereof, said exonuclease having the amino acid sequence of SEQ ID No. 1 or an amino acid sequence which is at least about 50% identical thereto, wherein said exonuclease or enzymatically active fragment thereof (i) is substantially irreversibly inactivated by heating at a temperature of about 55° C. for 10 minutes in a buffer consisting of 10 mM Tris-HCI, pH 8.5 at 25° C., 50 mM KCI and 5 mM MgCI2; (ii) is substantially specific for single stranded DNA; and (iii) has a 3?-5? exonuclease activity. The invention further provides a method of removing single stranded DNA from a sample, a method of nucleic acid amplification, a method of reverse transcription and a method of nucleic acid sequence analysis in which the exonuclease or enzymatically active fragment thereof is used.

Abstract: The invention provides an exonuclease or an enzymatically active fragment thereof, said exonuclease having the amino acid sequence of SEQ ID No. 1 or an amino acid sequence which is at least about 50% identical thereto, wherein said exonuclease or enzymatically active fragment thereof (i) is substantially irreversibly inactivated by heating at a temperature of about 55° C. for 10 minutes in a buffer consisting of 10 mM Tris-HCl, pH 8.5 at 25° C., 50 mM KCl and 5 mM MgCl2; (ii) is substantially specific for single stranded DNA; and (iii) has a 3?-5? exonuclease activity. The invention further provides a method of removing single stranded DNA from a sample, a method of nucleic acid amplification, a method of reverse transcription and a method of nucleic acid sequence analysis in which the exonuclease or enzymatically active fragment thereof is used.

Abstract: The present invention provides an endonuclease I or enzymatically active fragment thereof wherein said endonuclease I has the sequence of SEQ ID No. 4 or a sequence which is at least 70% identical thereto and wherein the amino acid residue which is immediately N-terminal of the FYCGC pentapeptide motif has been substituted with a residue which is negatively charged as well as nucleic acid molecules encoding these enzymes and methods of removing contaminating polynucleotides from a sample using these enzyme.

Abstract: The present invention provides an endonuclease I or enzymatically active fragment thereof wherein said endonuclease I has the sequence of SEQ ID No. 4 or a sequence which is at least 70% identical thereto and wherein the amino acid residue which is immediately N-terminal of the FYCGC pentapeptide motif has been substituted with a residue which is negatively charged as well as nucleic acid molecules encoding these enzymes and methods of removing contaminating polynucleotides from a sample using these enzyme.

Abstract: The present invention provides an endonuclease I or enzymatically active fragment thereof wherein said endonuclease I has the sequence of SEQ ID No. 4 or a sequence which is at least 70% identical thereto and wherein the amino acid residue which is immediately N-terminal of the FYCGC pentapeptide motif has been substituted with a residue which is negatively charged as well as nucleic acid molecules encoding these enzymes and methods of removing contaminating polynucleotides from a sample using these enzyme.

Abstract: The invention provides methods of removing nucleic acid contamination from reverse transcription reactions and hot-start PCR, wherein said hot-start PCR is a barrier hot-start PCR set up and/or involves a hot-start DNA polymerase, which methods comprise use of a DNase that is substantially irreversibly inactivated by heating at a temperature of about 50° C. for 5 minutes, and that is substantially specific for double stranded DNA. The invention further provides a DNase that is substantially irreversibly inactivated by heating at a temperature of about 50° C. for 5 minutes, and that is substantially specific for double stranded DNA, nucleic acids encoding said DNase and kits or compositions comprising said DNase or said nucleic acid.

Abstract: The present invention provides an endonuclease I or enzymatically active fragment thereof wherein said endonuclease I has the sequence of SEQ ID No. 4 or a sequence which is at least 70% identical thereto and wherein the amino acid residue which is immediately N-terminal of the FYCGC pentapeptide motif has been substituted with a residue which is negatively charged as well as nucleic acid molecules encoding these enzymes and methods of removing contaminating polynucleotides from a sample using these enzyme.

Abstract: The invention provides methods of removing nucleic acid contamination from reverse transcription reactions and hot-start PCR, wherein said hot-start PCR is a barrier hot-start PCR set up and/or involves a hot-start DNA polymerase, which methods comprise use of a DNase that is substantially irreversibly inactivated by heating at a temperature of about 50° C. for 5 minutes, and that is substantially specific for double stranded DNA. The invention further provides a DNase that is substantially irreversibly inactivated by heating at a temperature of about 50° C. for 5 minutes, and that is substantially specific for double stranded DNA, nucleic acids encoding said DNase and kits or compositions comprising said DNase or said nucleic acid.

Abstract: The invention provides methods of removing nucleic acid contamination from reverse transcription reactions and hot-start PCR, wherein said hot-start PCR is a barrier hot-start PCR set up and/or involves a hot-start DNA polymerase, which methods comprise use of a DNase that is substantially irreversibly inactivated by heating at a temperature of about 50° C. for 5 minutes, and that is substantially specific for double stranded DNA. The invention further provides a DNase that is substantially irreversibly inactivated by heating at a temperature of about 50° C. for 5 minutes, and that is substantially specific for double stranded DNA, nucleic acids encoding said DNase and kits or compositions comprising said DNase or said nucleic acid.

Abstract: The invention provides methods of removing nucleic acid contamination from reverse transcription reactions and hot-start PCR, wherein said hot-start PCR is a barrier hot-start PCR set up and/or involves a hot-start DNA polymerase, which methods comprise use of a DNase that is substantially irreversibly inactivated by heating at a temperature of about 50° C. for 5 minutes, and that is substantially specific for double stranded DNA. The invention further provides a DNase that is substantially irreversibly inactivated by heating at a temperature of about 50° C. for 5 minutes, and that is substantially specific for double stranded DNA, nucleic acids encoding said DNase and kits or compositions comprising said DNase or said nucleic acid.