| CPC C12Q 1/6816 (2013.01) [C12Q 1/6806 (2013.01); C12Q 1/6851 (2013.01); C12Q 1/6853 (2013.01); C12Q 1/6876 (2013.01); G16B 40/10 (2019.02); C12Q 2525/161 (2013.01); C12Q 2525/179 (2013.01); C12Q 2537/143 (2013.01); C12Q 2545/114 (2013.01); C12Q 2600/166 (2013.01)] | 20 Claims |
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1. A method for determination of abundance of a nucleic acid target from a sample including cell-free DNA (cfDNA), the method comprising:
generating a first set of oligonucleotides comprising different subsets of oligonucleotides, wherein each subset of the different subsets of oligonucleotides comprises oligonucleotides comprising:
a dilution tag sequence associated with the subset of the different subsets of oligonucleotides, wherein the dilution tag sequence is associated with a relative concentration profile indicating different relative concentrations of different dilution tag sequence combinations, and
a target-specific region complementary to a target sequence of the nucleic acid target;
generating a second set of oligonucleotides comprising dilution tag sequence-associated regions complementary to nucleotide sequences of the dilution tag sequences of the first set of oligonucleotides;
performing a labeling process with the first set of oligonucleotides and the cfDNA from the sample, thereby generating a dilution-tagged mixture comprising different subsets of the dilution-tagged nucleic acid target identified by the different dilution tag sequence combinations at different relative concentrations indicated by the relative concentration profile;
performing an amplification process with the second set of oligonucleotides and the dilution-tagged mixture;
modifying concentrations of the different subsets of the amplified dilution-tagged mixture to generate a modified dilution-tagged mixture; and
determining an abundance of the nucleic acid target using the modified dilution-tagged mixture and the relative concentration profile associated with the different dilution tag sequence combinations.
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