Large-scale insertional mutagenesis of a coleopteran stored grain pest, the red flour beetle Tribolium castaneum, identifies embryonic lethal mutations and enhancer traps

Table 5 Test for lethality and sterility

First round of F₃ single-pair matings (SPM)		Second round of F ₃ single-pair matings (SPM)
Offspring	Phenotype	Offspring	Phenotype
At least one SPM indicates homozygosity	Viable	At least one SPM (in total) indicates homozygosity	Viable
All informative* SPM indicate heterozygosity of both parents	Potentially lethal	All informative^* SPM indicate heterozygosity of both parents	Lethal
At least two SPM without any offspring but with living parents	Potentially sterile	Unable to find at least four SPM (in total) without any offspring but with living parents (method 1) OR unable to identify either a fertile homozygous female or a fertile homozygous male (method 2)	Sterile

After the first round of single-pair matings (SPM), all viable insertions were discarded (unless an enhancer trap was detected). All potentially lethal and potentially sterile lines were retested in a second round of single-pair matings.
*A single-pair mating is uninformative if it produces no offspring and one or both parents are dead, or less than 20 offspring are present and all of them are GFP-positive (see Table 4 and Methods)

ISSN: 1741-7007