SMAdd-seq of saccharomyces cerevisiae
The DNA/nuclei/spheroplast was placed on ice. 10 uL of a 2mg/ml angelicin* stock (0/20/500uM) (SIGMA-A0956) were added to each of the wells, the plate was swirled a few times to mix the angelicin and incubated in the dark on ice for 5 minutes. While ensuring the culture plate remained nested in ice, the plate was exposed to 365nm UVA light (Stratagene UV Stratalinker 2400, power 5.0) for 5 minutes followed by a 5-minute incubation in ice. This UV exposure process was repeated a further 6 times. Contents of both wells were pooled into a fresh low-adhesion tube (EPPENDORF-022431021) and both wells were washed with 100 uL of ice-cold angelicin modification buffer and added to the same low-adhesion tube to maximize nuclei retrieval. High molecular weight DNA was purified using the NEB Monarch HMW DNA Extraction kit for tissue (T3060L)