Extracted DNA was quantified with a Qubit 3.0 fluorometer and a DNA dsDNA HS Assay Kit (Life Technologies, Carlsbad, CA), as well as on the 2100 Bioanalyzer with High Sensitivity DNA Chips (Agilent Technologies, Santa Clara, CA) for assessment of sample purity, concentration, and fragment size distribution according to the manufacturer’s instructions. The average fragment size was determined with the Agilent 2100 Bioanalyzer Expert software, and calculated across the first three peaks 75–675 bp corresponding to the length of nucleosomal footprints and linkers derived from apoptotic cells (Supplementary Figure S1 ). The final plasma cfDNA concentrations were calculated by adjusting for the initial plasma and final elution volumes, and quantified with a Qubit 3.0 for a subset of patients (Supplementary Table S1 ). Assessment of cfDNA fragment size and concentration was performed without prior knowledge of clinical data. Average cfDNA fragment size was not available for mCRPC patients, since samples were not available for analysis on the 2100 Bioanalyzer.
Dna dsdna hs assay kit
Manufactured by Thermo Fisher Scientific
The DNA dsDNA HS Assay Kit is a fluorescence-based detection system designed to quantify double-stranded DNA (dsDNA) in small sample volumes. The kit provides a sensitive and accurate method for measuring dsDNA concentrations.
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2 protocols using dna dsdna hs assay kit
1
Quantification and Characterization of Circulating Cell-Free DNA
2
Quantification and Analysis of Plasma cfDNA
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Extracted DNA was quantified with a Qubit 3.0 Fluorometer and a DNA dsDNA HS Assay Kit (Life Technologies, Carlsbad, CA), as well as on the 2100 Bioanalyzer with High Sensitivity DNA Chips (Agilent Technologies, Santa Clara, CA) for assessment of sample purity, concentration, and fragment size distribution according to the manufacturer's instructions. The average fragment size was determined with the Agilent 2100 Bioanalyzer Expert software, and calculated across the first three peaks 75-675bp corresponding to the length of nucleosomal footprints and linkers derived from apoptotic cells (Supplementary Figure 1). The final plasma cfDNA concentrations were calculated by adjusting for the initial plasma and final elution volumes, and quantified with a Qubit 3.0 for a subset of patients (Supplementary Table 1). Assessment of cfDNA fragment size and concentration was performed without prior knowledge of clinical data. Average cfDNA fragment size was not available for mCRPC patients, since samples were not run on the 2100 Bioanalyzer.
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