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. 2021 Aug 18;112(10):4393–4403. doi: 10.1111/cas.15084

FIGURE 2.

FIGURE 2

Validation of cDNA synthesis and cfRNA extraction. A, Agarose gel electrophoresis of PCR amplicons of EML4‐ALK and KIF5B‐RET gene‐specific cDNA fragments from a standard RNA Mix. Estimated size bands were detected in all samples with indicated copy numbers of fusion genes. B, Real‐time PCR of GAPDH mRNA expression. Total cfRNA was extracted from plasma using the MagMAX Cell‐Free Total Nucleic Acid Isolation Kit (Kit A) or the Quick‐cfRNA Serum and Plasma Kit (Kit B). As a control, total RNA of H2228 cells was extracted using the RNeasy Mini Kit. cfRNA and RNA of H2288 cells were converted to cDNA using the GenNext RamDA‐seq Single Cell Kit. Real‐time PCR was performed using the Power SYBR Green Master Mix on the 7500HT Fast Real‐Time PCR System. Fold change of GAPDH mRNA expression was calculated by comparison with the value from 10 pg H2228 mRNA