DNDi6148 Sample 12 Lane 3
we used a set of twenty-six degenerate single-stranded oligodeoxynucleotides (ssODNs, Thermo Fisher Scientific), as described [17]. Briefly, we used 40 mg of each ssODNs in 10 ml of 10 mM Tris-Cl, pH 8.5, mixed with 5 million T. brucei inducible MSH2 RNAi cells in 100 ml transfection buffer. MSH2 knockdown was induced 24 h prior to transfection by the addition of 1 g/mL tetracycline (Sigma). Each ssODN was transfected individually and the cells were then pooled in 150 ml of medium to generate the library.