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US 2020/0399680 A1Published application
Nucleotide sequence exclusion enrichment by droplet sorting (needls)
AssigneeSamplix Aps
InventorsMarie Just Mikkelsen, Thomas Kvist
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Abstract
The present invention pertains to an in vitro method in which a targeted DNA molecule containing a DNA sequence of interest is enriched by a) general amplification of DNA molecules in a multiple of droplets each containing less than 0.5 target DNA molecule on average (404), b) specific detection of the target DNA molecule in each of the droplets (405), and c) physically selecting droplets containing target DNA molecules (406).
Claims15
1-15. (canceled)
An in vitro method for enriching for one or more target DNA molecules from a sample of mixed DNA molecules comprising the steps of:
- (a) providing a liquid sample of mixed DNA molecules comprising non-target DNA molecules and one or more specific target DNA molecules and reagents for general amplification of DNA (401);
- (b) formation of a multiple of liquid droplets each comprising mixed DNA molecules from said liquid sample (403);
- (c) specific detection of said one or more target DNA molecules in droplets containing at least one of said target DNA molecules (405);
- (d) physically selecting droplets containing at least one of said target DNA molecules (406), wherein the frequency of the target DNA molecule compared to its frequency in the sample of mixed DNA molecules in step (a) is increased between 0.1×(number of droplets without target DNA)×(total number of DNA containing droplets)−1 and 10×(number of droplets without target DNA)×(total number of DNA containing droplets)−1;
- and
- (e) general amplification of said non-target and target DNA molecules in the selected droplets.
3 more elements
Dependencies are inferred from the claim text.