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- Ahmadian, Afshin, et al.
(författare)
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Method for amplification
- 2005
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Patent (populärvet., debatt m.m.)abstract
- The invention refers to a method for multiplex amplification of at least one specific nucleic acid locus, comprising the steps of: providing at least one oligonucleotide probe pair that is designed so that the first and second probe of the pair anneal to a specific nucleic acid locus on a target molecule, in which pair the first probe has an extendable 3'-end, and a second probe has a 5'-end that is directly or indirectly labelled with a phosphate group; providing a target molecule comprising at least one specific nucleic acid locus; allowing the probe pair to anneal to the target molecule; allowing the 3'-end of the first probe to extend by influence of polymerase by adding a set of three different dNTPs; ligating the 3'-end of the extended first probe to the 5'-end of the second probe. Hereby, a method is provided which allows a high specificity for simultaneous amplification of several loci. Further, the invention involves a kit for use in the method of the invention.
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| 4. |
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| 5. |
- Lundeberg, Joakim, 1963-, et al.
(författare)
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Chemical method for the analysis of DNA sequences
- 1993
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Patent (populärvet., debatt m.m.)abstract
- The invention provides a method of identification of the base in a target position in a DNA sequence wherein sample DNA is subjected to amplification; the amplified DNA is immobilised and then subjected to strand separation, the non-immobilised strand being removed and an extension primer, which hybridises to the immobilised DNA immediately adjacent to the target position, is provided; each of four aliquots of the immobilised single stranded DNA is then subjected to a polymerase reaction in the presence of a dideoxynucleotide, each aliquot using a different dideoxynucleotide whereby only the dideoxynucleotide whereby only the dideoxynucleotide complementary to the base in the target position becomes incorpored; the four aliquots are then subjected to extension in the presence of all four deoxynucleotides, whereby in each aliquot the DNA which has not reacted with the dideoxynucleotide is extended to form double stranded DNA while the dideoxy-blocked DNA remains as non-extended DNA; followed by identification of the double stranded and/or non-extended DNA to indicate which dideoxynucleotide was incorporated and hence which base was present in the target position.
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| 6. |
- Lundeberg, Joakim, 1963-, et al.
(författare)
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Method for quantifying nucleic acid using multiple competitor nucleic acids
- 1997
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Patent (populärvet., debatt m.m.)abstract
- A method of determining the amount of target nucleic acid in a sample which comprises the steps of: a) adding to said sample a known amount of at least two different competitor nucleic acids at different concentrations, which have at least a portion of the sequence in common with the target nucleic acid, said common sequence comprising a binding site for a complementary primer sequence, b) co-amplifying the target nucleic acid and competitor nucleic acids in the sample by an in vitro amplification reaction using at least one primer, wherein at least one of said primers comprises a region complementary to said common sequence and the amplification products carry a label or means for attaching a label, c) separation of the amplification products, d) assessing the amount of label associated with the amplification products and e); comparison of the amount of label associated with each of the amplified target nucleic acid and amplified competitor nucleic acids to assess the amount of target nucleic acid in said sample, a method of diagnosis using the assay and kits for performing the same.
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| 7. |
- Lundeberg, Joakim, 1963-, et al.
(författare)
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Method of assessing the amount of nucleic acid in a sample
- 1999
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Patent (populärvet., debatt m.m.)abstract
- The present invention provides a method of assessing the amount of target nucleic acid in a sample which comprises co-amplifying the target nucleic acid and at least one competitor nucleic acid molecule wherein each competitor molecule contains a unique discriminatory sequence, and determining the relative amounts of the respective amplicons, characterised in that determination is achieved by detecting a primer extension reaction using each said amplicon as template and a kit for use in such a method.
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