Abstract: The present invention relates to melting analysis based methods for detecting the presence of a variant sequence in a target nucleic acid sequence comprising nucleotides of interest, in particular to detect microsatellite instability. The methods employ probes as reporter oligonucleotides with fluorophore and quencher and wherein the nucleotide sequence comprises nucleotides with hydrophobic intercalating residues. Also disclosed are methods for determining efficacy of a drug and for predicting the presence of a clinical disorder in an individual, as well as reporter oligonucleotides and kits for performing the methods.
Type:
Application
Filed:
May 13, 2020
Publication date:
June 30, 2022
Applicant:
PENTABASE APS
Inventors:
Kamilla Kolding Bendixen, Sarah Kronborg Eriksen, Emeli Elisabeth Hansen, Ulf Bech Christensen, Rasmus Koefoed Tersen, Cesilie Lind Madsen
Abstract: Provided herein are specially modified blocking nucleotides allowing for the sensitive detection of low copies of variant sequences, while significantly reducing signals from non-variant sequences that are similar but not identical to the variant sequence. These nucleotides can be used to detect rare variants in a sample mixture, as described in the present methods.
Abstract: Provided herein are specially modified blocking nucleotides allowing for the sensitive detection of low copies of variant sequences, while significantly reducing signals from non-variant sequences that are similar but not identical to the variant sequence. These nucleotides can be used to detect rare variants in a sample mixture, as described in the present methods.
Abstract: The invention relates to novel oligonucleotides comprising a signalling pair and at least two hydrophobic nucleotides. The oligonucleotide analogues are useful for detecting the status of nucleic acid sequences, such as presence, expression, methylation and/or mutation, in particular single point mutations and other sequences where the variation between the correct target and other targets may vary in as little as one nucleotide. The invention also relates to new ways of detecting sequence differences and optimizing conditions by using oligonucleotide analogues and readily available instruments. In particular the invention relates to specifically detecting quantity of a target nucleic acids or detecting one sequence over others that may vary in as little as one nucleotide using oligonucleotides or oligonucleotide analogues comprising a signalling pair and at least two hydrophobic nucleotides, such as a nucleotide analogue comprising an intercalator.
Abstract: The invention relates to novel oligonucleotides comprising a signalling pair and at least hydrophobic nucleotide. The oligonucleotide analogues are useful for detecting the status of nucleic acid sequences, such as presence, expression, methylation and/or mutation, in particular single point mutations and other sequences where the variation between the correct target and other targets may vary in as little as one nucleotide. The invention also relates to new ways of detecting sequence differences and optimizing conditions by using oligonucleotide analogues and readily available instruments. In particular the invention relates to specifically detecting quantity of a target nucleic acids or detecting one sequence over others that may vary in as little as one nucleotide using oligonucleotides or oligonucleotide analogues comprising a signalling pair and at least one hydrophobic nucleotide, such as a nucleotide analogue comprising an intercalator.