Phen-DC3 Trifluoromethanesulfonate (Phen-DC3 Triflate)

Cell experiment:

Briefly, untreated WT cells and pif1Δ cells from a fresh patch of cells get from a single colony bearing the parental allele size are diluted in 5 mL of YPD (2×105 cells/mL), grown for 8 generations at 30°C with shaking, and spreaded as single colony on YPD plates. To measure minisatellite instability upon Phen-DC3 Trifluoromethanesulfonate treatment, WT cells from a fresh patch on YPD are grown for 8 generations at 30°C in liquid SC containing Phen-DC3 Trifluoromethanesulfonate at 10 μM. Isolated colonies or pools of colonies are analyzed by Southern blot using the EcoRI digestion that cuts at each side of the minisatellite[2].

References:

[1]. Sanjay Kumar Bharti, et al. Specialization among Iron-Sulfur Cluster Helicases to Resolve G-quadruplex DNA Structures That Threaten Genomic Stability. J Biol Chem. 2013 Sep 27; 288(39): 28217–28229.
[2]. Aurèle Piazza, et al. Short loop length and high thermal stability determine genomic instability induced by G-quadruplex-forming minisatellites. EMBO J. 2015 Jun 12; 34(12): 1718–1734.