Hi All–
I need to separate wildtype worms from a mixed population of wildtype and Unc worms (with the nT1[unc(n754dm) let] balancer) in BULK. Does anyone know of a way to accomplish this feat?
Thanks!
Ash
Ashley B. Williams, Ph.D.
Molecular and Computational Biology
University of Southern California
Los Angeles, CA 90089
On thing that leaps to mind is to replace nT1 [n754] with nT1 [qIs51], and use either the COPAS biosorter or one of the published protocols to use a more standard FACS machine to sort C. elegans embryos.
It might also be possible to include a recessive drug resistance marker linked to nT1 in trans - avr-15, for example. This gets complicated and unwieldy, though.
I can imagine a serial enrichment of freely moving worms from Unc worms by chemotaxis assay or by settling in a water column (with modified viscosity, perhaps), but I don’t know of a protocol, and suspect the “in BULK” wouldn’t happen.
My recollection is that n754 is supposed to be the same as deg-3(u662), which doesn’t seem to be annotated anyplace I can readily find (the Treinin paper seems to agree with this, saying “The u662 mutation is the dominant Unc mutation associated with the DnT1 balancer chromosome”, but doesn’t name n754). So if you can find a good drug-hypersensitivity phenotype for deg-3(u662), that might do it for you.