[Zbrafish] Low efficacy of splice-blocking morpholino

[weibinz from umich.edu]

Hi,
I am testing two splicing -blocking morpholino rencently. I found the 
efficacy of blocking splicing is not high enough. I always detected the 
normaly processed mRNA in the embryos injected with morpholino, in the 
presence of the abberently processed mRNA due to the morpholino. My 
injected amount of morpholino is estimated to be about 16ng per zebrafish 
embryo, which I think has hit the upper limit and some embryos( about 10%) 
already showed massive necrosis. I used either Danieau's or water to dilute 
the morpholino to 0.5mM concentration and inject 2-4 nl per embryo. I am 
wondering whether anybody could share with me some tips to increase the 
efficacy.

Thanks a lot.

weibin