A strain I’m trying to inject is apparently very ‘sensitive’ to the Halocarbon oil (Sigma H8898) - placing worms in a drop of the oil for a few minutes results in worms that cannot be recovered when retrieved onto NGM with M9 in the usual way. N2s are fine. If I get through each injection as quickly as possible then I can get approx 10-20% recovery. Is there a substitute for this oil and/or are different purities available?
Hi,
a discussion way way back considered the merits of different halocarbon oil suppliers and alternatives (see post and followup);
http://www.bio.net/bionet/mm/celegans/1997-October/001492.html
this maybe a good starting point as I guess it will be suck it and see in terms of finding something that is right for your mutant?
Steve
Hmm, have you tried to use recovery buffer instead of M9?
Recipe: 5mm HEPES pH 7.2, 3 mM CaCl2, 3 mM MgCl2, 66 mM NaCl, 2.4 mM KCl, 4% Glucose (w/v) url=http://www.wormbook.org/chapters/www_transformationmicroinjection/transformationmicroinjection.html[/url]
yes, good point…skipped over the mention of M9…are you one…twoing the worms with recovery buffer then gradually into M9, especially if your strain is osmotically sensitive?
Steve
No I’ve just been recovering ‘once’ in M9. I’ll try more extensive washing with ‘recovery buffer’. Thanks.