| CPC C12Q 1/6806 (2013.01) [C12Q 1/6886 (2013.01); G16B 25/00 (2019.02); G16B 25/20 (2019.02); G16B 30/00 (2019.02); C12Q 2600/166 (2013.01)] | 20 Claims |
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1. A method of validating a patient-specific oligo, the method comprising:
extracting DNA or RNA from a patient sample to produce a DNA or RNA sample;
introducing a plurality of sets of spike-in sequences into the DNA or RNA sample to produce a spiked sample, wherein each spike-in sequence is an oligonucleotide sequence unrelated to any sequence in the sample from the patient, wherein each set of spike-in sequences comprises a plurality of spike-in sequences with a reference sequence unique to a set of spike-in sequences in the plurality of sets of spike-in sequences, wherein all spike-in sequences in a set of spike-in sequences comprise a sequence complementary to a synthesis-control oligo, and wherein the plurality of sets of spike-in sequences comprises spike-in sequences of known concentrations;
assaying the spiked sample with synthesis-control oligos comprising oligos complementary to the spike-in sequences for each set of spike-in sequences and patient-specific oligos comprising oligos complementary to sequences in the DNA or RNA sample to produce a read out of the counts of the plurality of sets of spike-in sequences and detect the patient-specific RNA or DNA; and
determining the correspondence between the counts from the sets of spike-in sequences with the known concentrations of the spike-in sequences to validate a batch synthesis of the patient-specific oligos, wherein the batch synthesis of the patient-specific oligos is validated if the the counts from the sets of spike-in sequences correspond to the known concentrations from the sets of spike-in sequences.
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