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Amplified ccfDNA for Assay Development, Validation, and Proficiency Testing

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"Amplified ccfDNA for Assay Development, Validation, and Proficiency Testing"

Circulating cell free DNA is the analyte for Non Invasive Prenatal Testing assays and circulating tumor DNA assays.

Liquid biopsies are now carried out in many labs and the combined number is now around a million tests per year in the US and these assays continue to become more sensitive and specific by becoming tuned to the nuances of ccfDNA. Since ccfDNA may be present in plasma at only 5 ng/ml, and since 50 ng or more may be the preferred input for some assays, native ccfDNA can usually not be obtained at a sufficient amount to be useful as a reference, proficiency testing, or control material. We developed a method to amplify ccfDNA so that nanograms of input can generate micrograms of output that would be sufficient for hundreds to thousands of tests. Here we analyzed how amplified ccfDNA performs in relation to matched native DNA and whether it sufficiently reproduces the nuances of ccfDNA

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