Magmax cell free dna isolation kit mm

Ten milliliters of peripheral blood samples were obtained immediately before ultrasound-guided core needle biopsy. Plasma from Streck BCT tubes was prepared within 2 h after blood collection and stored at − 20 °C in the clinic until shipment to the research laboratory. cfDNA was isolated from 5 ml of plasma with a MagMAX Cell-Free DNA Isolation Kit (MM; Applied Biosystems, Thermo Fisher Scientific, Foster City, CA, USA) and then eluted in 60 µl of elution buffer according to the manufacturer’s protocol. cfDNA was quantified using a QuantiFluor dsDNA System and GloMax Discover Microplate Reader (Promega, Madison, WI, USA). The distribution of fragment lengths was checked by electrophoresis on an Agilent 2100 Bioanalyzer with a High Sensitivity Large Fragment 50 kb DNA Kit (Agilent, Technologies Inc., Santa Clara, CA, USA). An NEBNext Ultra II DNA Library Prep kit (New England Biolabs, UK; E7645) was used for cfDNA whole-genome library preparation. Higher-pass whole-genome sequencing was started with 10 ng of cfDNA input (median of 5 ng). Finally, 32 libraries were pooled and sequenced using 150 bp pair-end run reads and 8 bp dual-indices on an Illumina NovaSeq machine (Illumina, San Diego, CA, USA), producing cfDNA whole-genome sequences for each subject.