I need to generate isofemale lines of Drosophila melanogaster but the last time we initiated a set of such lines, most of the lines crashed possibly due to inbreeding depression. I am aware that inbreeding depression is something I will have to deal with when it comes to generating such lines but if you have any experience with isofemale lines, can you suggest me any useful tips or strategies I can use to prevent (as much as possible) lines from crashing?
are u trying to isogenize any particular mutant or wild type strain?
at which generation does the line crash ? as in F1 or F5 for instance ?
1) Start with loads of sib crosses because you will lose lots of them
2) after a couple of generations when you don't want to lose the line as you have invested in them, run 2 or 3 sib crosses per line, so that if any one sib pair fails to produce offspring, you have a backup.
3) keep the parents for a while (ie don't discard after a week of laying). If you let the adults lay, then move them to a colder incubator, they can survive for till you know whether their offspring are producing offspring. If their offspring fail to produce, you can get more from the parents if you have kept them alive
4) good husbandry is key- larvae do badly when there are few of them, so be very careful with your recipes and keeping fungi and bacteria away
Good luck!
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