Abstract: The present invention relates to a polymerase enzyme with improved ability to incorporate reversibly terminating nucleotides. The enzyme comprising the following mutations in the motif A region (SGS). It relates to a polymerase enzyme according to SEQ ID NO. 1 or any polymerase that shares at least 70% amino acid sequence identity thereto, comprising a mutation selected from the group of (i) at position 412 of SEQ ID NO. 1: serine (S) (L412S) and/or, (ii) at position 413 of SEQ ID NO. 1: glycine (G) (Y413G) and/or (iii) at position 414 of SEQ ID NO. 1: serine (S) (P414S), wherein the enzyme has little or no 3?-5? exonuclease activity.

Abstract: A method for assessing the status of degradation and/or integrity of one or more nucleic acids in a sample, which includes amplifying at least two overlapping regions within at least one locus and detecting the amount of the amplification products using at least two probes, where one probe binds to an overlapping region and the other probe binds to a non-overlapping region. Also, a method of designing primers and/or probes for amplifying at least two overlapping regions within at least one single copy locus or multicopy locus. Also a primer and a primer pair for amplifying at least two overlapping regions from one nucleic acid template which is a multicopy locus present in loci in the human genome. Also a kit including primers and/or probes for assessing the status of degradation and/or integrity of one or more nucleic acids in a sample.

Abstract: Oligonucleotide primers and probes for quantifying and/or detecting human male DNA in a forensic sample. The oligonucleotides are useful for amplifying a multicopy locus within the human Y-chromosome (MCL-Y) that shares at least 80% sequence identity to a sequence according to SEQ ID NO. 3 over a stretch of at least 60 base pairs. The oligonucleotides hybridize under stringent conditions to a nucleic acid having at least one sequence selected from the group consisting of SEQ ID NO. 3 to SEQ ID NO. 11 and/or SEQ ID NO. 17 to SEQ ID NO. 25. Kits including the oligonucleotide primers, probes and/or primer pairs and reagents for performing an amplification reaction on DNA recovered from a forensic sample are also disclosed.

Abstract: The invention is in the field of regulation of enzymatic activity in nucleic acid modifying reactions. It describes a method of regulating enzymatic activity by adding chelating agents to the reaction composition and exploits the fact that both the binding of divalent cations to these chelating agents and the pH of commonly used buffers is temperature dependent. PCR experiments that are hampered by non-specific side products can be regulated such that the target sequence is amplified in a more specific manner.

Abstract: The present invention provides methods and kits for virus inactivation by virus-deactivating substances and direct amplification-based detection of target nucleic acids and viruses in biological samples without prior target purification.

Abstract: The present invention provides method and kits for amplification based detection of target nucleic acids and pathogens using crude biological samples without prior target purification.

Abstract: The present invention provides method and kits for amplification based detection of target nucleic acids and pathogens using crude biological samples without prior target purification.

Abstract: A kit for use in assessing the status of nucleic acid degradation and/or the integrity of one or more nucleic acids in a sample by amplifying at least two overlapping regions within at least one locus and detecting the amount of the at least two amplification products. The kit includes a primer and at least two probes that bind under stringent conditions to a sequence that shares at least 80% sequence identity to a sequence selected from the group of sequences consisting of SEQ ID NO. 6 to SEQ ID NO. 47 over a stretch of 80 base pairs, or to a reverse complement thereof. One of the at least two probes binds to one of the at least two overlapping regions and the other of the at least two probes binds to a non-overlapping region.

Abstract: The invention is in the field of regulation of enzymatic activity in nucleic acid modifying reactions. It describes a method of regulating enzymatic activity by adding chelating agents to the reaction composition and exploits the fact that both the binding of divalent cations to these chelating agents and the pH of commonly used buffers is temperature dependent. PCR experiments that are hampered by non-specific side products can be regulated such that the target sequence is amplified in a more specific manner.

Abstract: An oligonucleotide suitable for use as a detection probe in a method for evaluating the amplification efficiency, the presence of inhibitors, degradation and/or for performing a quantification analysis of a target nucleic acid in a real-time amplification reaction. The oligonucleotide probe is preferably SEQ ID NO. 4, a reverse complement of SEQ ID NO. 4, an oligonucleotide that shares 95% sequence identity with SEQ ID NO. 4, or an oligonucleotide that shares 95% sequence identity with a reverse complement of SEQ ID NO. 4. Also a kit including an internal nucleic acid control template, a set of primers for amplifying the internal nucleic acid control template, and the oligonucleotide probe.

Abstract: The present invention relates to a polymerase enzyme with improved ability to incorporate reversibly terminating nucleotides. The enzyme comprising the following mutations in the motif A region (SGS). It relates to a polymerase enzyme according to SEQ ID NO. 1 or any polymerase that shares at least 70% amino acid sequence identity thereto, comprising a mutation selected from the group of (i) at position 412 of SEQ ID NO. 1: serine (S) (L412S) and/or, (ii) at position 413 of SEQ ID NO. 1: glycine (G) (Y413G) and/or (iii) at position 414 of SEQ ID NO. 1: serine (S) (P414S), wherein the enzyme has little or no 3?-5? exonuclease activity.

Abstract: PCR that allows the researchers to amplify a desired DNA requiring only tiny amounts of sample. Such amplification reactions are technically challenging and are often hampered by several practical issues such as the presence of PCR inhibitors, sample degradation and low quantities of said sample. The invention addresses these issues with a method for evaluating the amplification efficiency and/or the presence of inhibitors and/or degradation and/or performing a quantification of a nucleic acid in a real-time amplification reaction comprising: optionally amplifying in a reaction composition a first target nucleic acid using a first primer pair in a real-time amplification reaction, (i) amplifying in said reaction composition one or more second internal nucleic acid control templates (IC) with a length of between 50 and 2000 nucleotides, wherein the second nucleic acid has a sequence selected from the group of: i) SEQ ID NO.

Abstract: A kit for use in assessing the status of nucleic acid degradation and/or the integrity of one or more nucleic acids in a sample by amplifying at least two overlapping regions within at least one locus and detecting the amount of the at least two amplification products. The kit includes a primer and at least two probes that bind under stringent conditions to a sequence that shares at least 80% sequence identity to a sequence selected from the group of sequences consisting of SEQ ID NO. 6 to SEQ ID NO. 47 over a stretch of 80 base pairs, or to a reverse complement thereof. One of the at least two probes binds to one of the at least two overlapping regions and the other of the at least two probes binds to a non-overlapping region.

Abstract: Oligonucleotide primers and probes for quantifying and/or detecting human male DNA in a forensic sample. The oligonucleotides are useful for amplifying a multicopy locus within the human Y-chromosome (MCL-Y) that shares at least 80% sequence identity to a sequence according to SEQ ID NO. 3 over a stretch of at least 60 base pairs. The oligonucleotides hybridize under stringent conditions to a nucleic acid having at least one sequence selected from the group consisting of SEQ ID NO. 3 to SEQ ID NO. 11 and/or SEQ ID NO. 17 to SEQ ID NO. 25. Kits including the oligonucleotide primers, probes and/or primer pairs and reagents for performing an amplification reaction on DNA recovered from a forensic sample are also disclosed.

Abstract: The present invention relates to a method for assessing the status of nucleic acid degradation and/or integrity of one or more nucleic acids in a sample, comprising the steps amplifying at least two overlapping regions within at least one locus (e.g. by heminested or nested PCR), and detecting the amount of the at least two amplification products through the use of at least two probes, wherein one probe binds to the region of overlap and the at least one other probe binds to a non-overlapping region. The invention further relates to a method of designing primers and/or probes for amplifying at least two overlapping regions within at least one locus, wherein the locus that is amplified is a single copy locus (SCL) or multicopy locus (MLC). The invention also relates to a primer and a primer pair for amplifying at least two overlapping regions from one nucleic acid template which is a multicopy locus present in 21 loci in the human genome. These primers and probes may be in a kit.

Abstract: The present invention relates to a family B polymerase enzyme comprising the following mutations, a mutation in the motif A region, wherein a threonine is the second amino acid of the motif A region, wherein the motif A region is homologous to amino acids 408 to 410 in 9° N, a mutation leading to a reduction or elimination of the 3?-5? exonuclease activity if present in said family B polymerase, wherein said polymerase additionally comprises one or both of the following mutations in the motif A region, L408Y or L408F and/or P410S or P410G. It may be used in DNA sequencing and may be present in a kit.

Abstract: According to a first aspect of the present invention, a method is provided for detecting and/or quantifying male genomic DNA in a sample, wherein the method comprises the step of amplification of a multicopy locus within the human Y-chromosome (MCL-Y), wherein said locus shares at least 80% sequence identity to a sequence according to SEQ ID NO. 3 over a stretch of at least 60 base pairs (bp). A second aspect of the present invention relates to a primer or primer pair which hybridizes under stringent conditions to a sequence according to SEQ ID NO. 3 and/or any of 4 to 11. The invention also relates to a kit.

Abstract: The invention is in the field of regulation of enzymatic activity in nucleic acid modifying reactions. It describes a method of regulating enzymatic activity by adding chelating agents to the reaction composition and exploits the fact that both the binding of divalent cations to these chelating agents and the pH of commonly used buffers is temperature dependent. PCR experiments that are hampered by non-specific side products can be regulated such that the target sequence is amplified in a more specific manner.

Abstract: The present invention relates to a family B polymerase enzyme comprising the following mutations, a mutation in the motif A region, wherein a threonine is the second amino acid of the motif A region, wherein the motif A region is homologous to amino acids 408 to 410 in 9° N, a mutation leading to a reduction or elimination of the 3?-5? exonuclease activity if present in said family B polymerase, wherein said polymerase additionally comprises one or both of the following mutations in the motif A region, L408Y or L408F and/or P410S or P410G. It may be used in DNA sequencing and may be present in a kit.

Abstract: The invention is in the field of regulation of enzymatic activity in nucleic acid modifying reactions. It describes a method of regulating enzymatic activity by adding chelating agents to the reaction composition and exploits the fact that both the binding of divalent cations to these chelating agents and the pH of commonly used buffers is temperature dependent. PCR experiments that are hampered by non-specific side products can be regulated such that the target sequence is amplified in a more specific manner.