The human plasma samples were filtered (Minisart NML syringe filters, pore size: 0.8 μm; Sartorius) and diluted with 1X phosphate‐buffered saline without calcium and magnesium buffer (Gibco) at 1:3 (v/v) ratio and processed for EV isolation by a charge—based isolation method as described before (Notarangelo et al., 2020 (link), 2019 (link)). The diluted plasma was recovered and processed for cfDNA isolation by QIAmp Circulating Nucleic Acid kit (Qiagen) (Lee et al., 2018 (link)).
Mugoni V., Ciani Y., Quaini O., Tomasini S., Notarangelo M., Vannuccini F., Marinelli A., Leonardi E., Pontalti S., Martinelli A., Rossetto D., Pesce I., Mansy S.S., Barbareschi M., Ferro A., Caffo O., Attard G., Di Vizio D., D'Agostino V.G., Nardella C, & Demichelis F. (2023). Integrating extracellular vesicle and circulating cell‐free DNA analysis using a single plasma aliquot improves the detection of HER2 positivity in breast cancer patients. Journal of Extracellular Biology, 2(9), e108.
Pore size of Minisart NML syringe filters (0.8 μm)
Dilution ratio of plasma with 1X phosphate-buffered saline (1:3, v/v)
Use of phosphate-buffered saline without calcium and magnesium
positive controls
Not explicitly mentioned
negative controls
Not explicitly mentioned
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