Generation of ligation-ready dna amplicons
AssigneeQiagen GmbH
InventorsNan Fang, Dirk Loeffert
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Abstract
The invention is directed to novel methods, kits and uses to be employed for the generation of ligation-ready DNA amplicons of a target DNA by using 5′-phosphorylated primers.
Claims15
A method for generating ligation-ready DNA amplicons of a target DNA, comprising
- (i) contacting in a polymerase chain reaction (PCR) buffer said target DNA with at least one DNA polymerase, a dNTP mixture, and at least one PCR primer pair consisting of two target specific PCR primers, to obtain a reaction mixture,
- (ii) subjecting said reaction mixture to a PCR to generate a plurality of ligation-ready DNA amplicons of said target DNA,
- wherein at least one of said target specific PCR primers is 5′-phosphorylated.
A method for generating DNA-adaptor-ligated DNA amplicons of a target DNA, comprising
- (i) contacting in a polymerase chain reaction (PCR) buffer said target DNA with at least one DNA polymerase, a dNTP mixture, and at least one PCR primer pair consisting of two target specific PCR primers, to obtain a reaction mixture,
- (ii) subjecting said reaction mixture to a PCR to generate a plurality of ligation-ready DNA amplicons of said target DNA,
- (iii) isolating said plurality of ligation-ready DNA amplicons of said target DNA from said reaction mixture,
- (iv) ligating said ligation-ready DNA amplicons of said target DNA to at least one DNA adaptor molecule to generate DNA-adaptor-ligated DNA amplicons of said target DNA,
- wherein at least one of said target specific PCR primers is 5′-phosphorylated.
The method of claim 10, wherein said sequencing primer is a primer for next generation sequencing (NGS).
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A method for generating ligation-ready DNA amplicons of a target DNA, the method comprising amplifying the target DNA using at least one 5′-phosphorylated PCR primer.
Dependencies are inferred from the claim text.