Figure 2.

PspGI digestion reduced the total number of PCR screenings and allowed for screening with multiple primer pairs. DNA lysates from a library of mutagenized worms (approximately 100 000 genomes) were pooled and screened with 12 reactions. PCRs were performed with multiplexed slo-2 primers covering the N-terminus and the C-terminus. Two initial positives were detected (lanes 5 and 9). Only one positive (lane 5) verified a 1.7 kb product with triplicated PCRs, and yielded a slo-2 C-terminus deletion strain, slo-2(nf101). PCR products resolved by gel electrophoresis on a 2% agarose gel with a 1 kb DNA size marker (Gibco).