Methods and compositions for enrichment of amplification products
AssigneeAccuScan Sciences Inc.
InventorsLi WENG, Shengrong LIN, Lin Fung TANG
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Abstract
In some aspects, the present disclosure provides methods for enriching amplicons, or amplification products, comprising a concatemer of at least two or more copies of a target polynucleotide. In some embodiments, a method comprises sequencing the amplicons comprising at least two or more copies of a target polynucleotide. In some embodiments, the target polynucleotides comprise sequences resulting from chromosome rearrangement, including but not limited to point mutations, single nucleotide polymorphisms, insertions, deletions, and translocations including fusion genes. In some aspects, the present disclosure provides compositions and reaction mixtures useful in the described methods.
Claims20
A method of increasing a proportion of concatemers having at least two copies of a target polynucleotide generated by a rolling circle amplification, comprising:
- (a) providing a circular polynucleotide comprising a target polynucleotide;
- (b) subjecting an amplification reaction mixture to multiple cycles of rolling circle amplification to generate a plurality of amplification products comprising concatemers, wherein the amplification reaction mixture comprises (i) a polymerase having strand displacement activity, (ii) the circular polynucleotide, and (iii) primers;
- and wherein each cycle of the multiple cycles of rolling circle amplification comprises denaturation at a denaturing temperature, primer annealing at an annealing temperature, and primer elongation at an elongation temperature for a given elongation time period, to generate the plurality of amplification products;
- thereby increasing a proportion of concatemers having at least two copies of the target polynucleotide.
Dependencies are inferred from the claim text.