Mapping Mutations in C. elegans - Caenorhabditis Elegans: Molecular Genetics and Development - page 6

Fig. 1 Flowchart for characterization of newly isolated mutations. Overall progression and major
conclusions are diagrammed. See text for details and caveats.
WGS. Despite the large number of mutations induced by standard mutageneses
( Anderson, 1995; Flibotte et al., 2010; Sarin et al., 2010 ), if a particular gene is
found to be mutated in each of two independently isolated, non-complementing
mutants, then it is overwhelmingly likely that this is the gene of interest. In this
case, it is important to sequence non-mutant siblings from the same mutant screen to
rule out mutations that have occurred spontaneously during growth of the stock.
2. Assessment of Penetrance
Examine the full brood of at least three individual New hermaphrodites. To do
this, pick a single L4-stage New hermaphrodite onto each of three seeded plates.
After 24 h (at 20 ), transfer each hermaphrodite to a new seeded plate, and repeat the
transfer at 12 h intervals until the hermaphrodites are purged (have run out of
sperm). Examine each plate daily for the presence of unhatched eggs (indicating
embryonic lethality) or slow-growing/arrested larvae. If possible, score every animal
on the plate for the New phenotype.