I’m having trouble locating a simple daf-2/daf-16 double mutant. I want to work with the canonical deletions of each: daf-2(e1370)III and daf-16(mu86)I.
I can find many double mutant strains with these phenotypes, but they all contain additional constructs. I can find other daf-16 alleles such as daf-16(m26)I; daf-2(e1370)III.
(m26) appears to render daf-16 non-functional so it will do in a pinch, but I’m surprised to find so many strains with daf-16(mu86); daf-2(e1370) and various gfp constructs, without there being any double mutants of these canonical mutants without any additional constructs.
Has anyone else encountered this?
Order one of those strains, pick a few non-Roller progeny to plates, one per plate, and make sure they don’t throw Roller progeny. You then have the daf-16(mu86); daf-2(e1370) strain you desire. You can read more about the construction of those strains in this paper.
Ditto to what Hillel said.
Also, you may want to note that e1370 is not a deletion allele of daf-2. m65 is a likely null. The CGC does have a double daf-16(mgdf50); daf-2(m65).
Please note that daf-16(m26) may be a likely hypomorph; many splice site mutations are strong reduced function alleles but not null. Just because it is the reference allele doesn’t mean it is the best. I’d go with something else.
daf-2(e1370) is a moderate allele, non-null. Though in many cases daf-2(e1370) is much easier to work with compared to the m65 null.