Single-Cell Genome Amplification via WGA-X

Prior to single-cell genome amplification, cells were lysed by one freeze-thaw cycle at −80°C and then at 20°C. The amplification procedures followed the WGA-X protocol (53 (link)), except that the total volume was 5 µL instead of 10 µL. In brief, the WGA-X components are as follows: 0.2 U µL⁻¹ Equiphi29 polymerase (Thermo Fisher Scientific), 1 $\times$ Equiphi29 reaction buffer (Thermo Fisher Scientific), 10 mM dithiothreitol (Thermo Fisher Scientific), 40 mM Exo-Resistant Random Primer (Thermo Fisher Scientific), 0.4 mM dNTP (New England BioLabs), and 1 µM SYTO-13 (Thermo Fisher Scientific). The WGA-X reaction was carried out by using a CFX384 TouchTM Real-Time Detection System (Bio-Rad Laboratories) for 7 hours at 45°C and then inactivated by incubation at 75°C for 15 min.