DETECTION PROBE FOR INTERNAL AMPLIFICATION CONTROL AND KIT INCLUDING SAME

    公开(公告)号:US20220333176A1

    公开(公告)日:2022-10-20

    申请号:US17748928

    申请日:2022-05-19

    申请人: QIAGEN GmbH

    摘要: 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.

    INTERNAL AMPLIFICATION CONTROL
    4.
    发明申请

    公开(公告)号:US20200087720A1

    公开(公告)日:2020-03-19

    申请号:US16333143

    申请日:2017-09-15

    申请人: QIAGEN GmbH

    摘要: 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. 1 or a sequence that is differs by no more than 15% therefrom, ii) a reverse complement of SEQ ID NO. 1 or a sequence that is differs by no more than 15% therefrom, iii) a sequence or a reverse complement thereof comprising the final 18 to 30 3′-nucleotides of SEQ ID NO. 1 at its terminal 3′-end and the final 18 to 30 5′-nucleotides of SEQ ID NO. 1 at its terminal 5′-end, or terminal ends that differ by no more than 5% from SEQ ID NO. 1, wherein between these terminal 3′-ends and 5′-ends the nucleic may have any nucleotide sequence and is between about 30 and about 1950 nucleotides in length.

    METHOD FOR DETERMINING NUCLEIC ACID DEGRADATION IN A SAMPLE IN WHICH AT LEAST TWO OVERLAPPING AMPLICONS ARE PRODUCED AND TWO PROBES ARE USED IN THE METHOD

    公开(公告)号:US20190264256A1

    公开(公告)日:2019-08-29

    申请号:US16333043

    申请日:2017-09-15

    申请人: QIAGEN GmbH

    IPC分类号: C12Q1/6806 C12Q1/6851

    摘要: 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. Hence, the invention also relates to a kit for assessing the status of nucleic acid degradation and/or integrity of one or more nucleic acids in a sample.

    KIT FOR DETERMINING NUCLEIC ACID DEGRADATION

    公开(公告)号:US20220025451A1

    公开(公告)日:2022-01-27

    申请号:US17500204

    申请日:2021-10-13

    申请人: QIAGEN GmbH

    IPC分类号: C12Q1/6851

    摘要: 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.