Single hepatocytes from each subject were subjected to WGA using our modified procedure of low-temperature cell lysis and DNA denaturation followed by MDA as described (11). As positive and negative controls for WGA, we used 1 ng of human genomic DNA and DNA-free PBS solution, respectively. Resultant MDA products were purified using AMPureXP beads (Beckman Coulter), and the amplified DNA concentration was measured with the Qubit High Sensitivity dsDNA kit (Invitrogen Life Sciences). To verify sufficient and uniformly amplified single-cell MDA products, we performed the eight-target locus-dropout test as described previously (11). Selected confirmed samples (four single-cell MDA products per subject) were further subjected to library preparation and WGS.

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