Fig. 3

Analysis of PIK3CA T1035A mutation in CTCs, primary tumors, and lung metastases from BC-PDX model BCM-4888. a Whole genome amplification was conducted on single cells using Rubicon PicoPLEX® kit, followed by nested PCR using primers spanning the PIK3CA mutation. PCR products were run on an agarose gel to confirm the presence of an amplicon of the correct size. Images of the single cells picked and then used for the WGA are shown below the gel image (scale varies between cells). b Sanger sequencing was applied to the purified amplicons using internal primers to confirm presence of the T1035A mutation in the primary tumor cells, CTCs, and lung metastasis cells. A wild type reference derived from HeLa DNA was run in parallel. This is a representative image of the sequencing trace from the cells containing the mutated gene