fig3

Figure 3. Comparison of two RNA extraction columns (Norgen vs. Qiagen) for detecting the EML4-ALK translocation by dPCR. EV-enriched (fractions 9-11), LPP-enriched (15-17), and protein-enriched (19-21) fractions from H2228 were used. RNA was eluted in 40 µL (Norgen) or 14 µL (Qiagen). From each, 6.5 µL was used for cDNA synthesis. Each dot represents a well on the dPCR chip. Blue dots represent wells with at least one copy of the EML4-ALK translocation, (FAM fluorophore); red dots show detection of the endogenous PUM1 gene (VIC fluorophore). Yellow and green dots indicate wells negative or positive for both genes, respectively. Grey dots represent wells not assigned by the analysis software. Graphing software: QuantStudio 3D AnalysisSuite Cloud Software. EML4-ALK: Echinoderm microtubule-associated protein-like 4-anaplastic lymphoma kinase; cDNA: complementary DNA; dPCR: digital polymerase chain reaction; EV: extracellular vesicle; FAM: 6-carboxyfluorescein; LPP: large protein particle; PUM1: pumilio RNA-binding family member 1; RNA: ribonucleic acid.