Generation of tagged dna fragments
OwnerIoanna Andreou, Nan Fang +2inventors
InventorsIoanna Andreou, Nan Fang, Dirk Loeffert, Annika Piotrowski
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Abstract
The present invention is directed to novel methods, kits and uses to be employed for the generation of tagged DNA fragments of a target DNA and nucleic acid molecules associated therewith
Claims15
A method for generating tagged DNA fragments of a target DNA, comprising
- (i) contacting said target DNA with an integrase and at least one DNA adaptor molecule that comprises an integrase recognition site, to obtain a reaction mixture,
- (ii) incubating said reaction mixture under conditions wherein a 3′ processing of said at least one adaptor molecule and a strand transfer reaction is catalyzed by said integrase, wherein
- (a) said target DNA is fragmented to generate a plurality of target DNA fragments, and
- (b) said at least one DNA adaptor molecule is joined to at least one end of each of the plurality of said target DNA fragments,
- to generate a plurality of tagged DNA fragments of said target DNA.
A kit for generating tagged DNA fragments of a target DNA, comprising:
- (i) an integrase, and
- (ii) at least one DNA adaptor molecule comprising an integrase recognition site.
The kit of claim further comprising at least one PCR primer pair configured to add in a PCR reaction to said at least one DNA adaptor molecule a site for annealing an oligonucleotide, wherein said site for annealing an oligonucleotide is configured for annealing a PCR and/or sequencing primer.
A method of using an integrase for generating a library of tagged DNA fragments of a target DNA, preferably said library of tagged DNA fragments is a library to be used for DNA sequencing, preferably via next generation sequencing.
A nucleic acid molecule comprising a nucleotide sequence selected from the group consisting of SEQ ID NOs. 1 to 15.
Dependencies are inferred from the claim text.