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Fig. 3 | BMC Cancer

Fig. 3

From: A rapid and quantitative method to detect human circulating tumor cells in a preclinical animal model

Fig. 3

QPCR evaluation of human CTC numbers in peripheral mouse blood. a For the generation of absolute QPCR analysis, the mouse GUS PCR products were cloned into a TA vector, followed by gene sequencing, E. coli amplification, and plasmid purification. The molecular weight of the plasmid was calculated using the value of OD.260 and diluted into 108 to 102 copy number/μl. b For the specificity of GUS gene primers for human- and mouse-purified DNA (upper-left panel), the PCR products were evaluated by melting curve analysis after quantitative analysis for single product confirmation (lower-left panel). DNA from the mouse blood containing human CTCs was measured by applying both human and mouse GUS primer sets. DNA from the mice with PBS (no cells) was included as a control (upper-right panel). The PCR products from the mouse blood containing CTCs were evaluated by melting curve analysis after quantitative analysis for single product confirmation (lower-right panel). c The QPCR-calculated cell numbers for each mouse were compared with the known IV-injected breast cell numbers. The human GUS cell numbers were normalized with the corresponding mouse GUS results. The correlation between the human GUS copy numbers and the injected cell numbers are indicated as R2 values. Bar errors are represented by three independent experiments

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