DAPI staining - (Jul/16/2005 )

hi
Im trying to do MLO staing on plant sections. the protocol I'm using says to dissolve DAPI in phosphate buffer pH 7.4, at 1 microgram per ml but it doesn't dissolve properly. pls somebody tell me how to do it, thanks in advance
chandima

Hey,
DAPI kind of ppts out in the PBS buffer that I use also. It does not mix up when yu vortex it...so what I do is I make my stock concentration in say 75 ul and pipete it up and down using a pipete tip. Make sure that its is dispersed well and then take say 20 ul add it to 1 ml. again pipete it....then add it to say 9 ml of PBS.
so basically the idea is to pipete and not vortex.
good luck
p..

maybe try to first dissolve your DAPI in water at 1 mg/ml and after make your solution to use on cells at desired concentration in PBS