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[Zbrafish] Low efficacy of splice-blocking morpholino

[Zbrafish] Low efficacy of splice-blocking morpholino - Zebrafish Forum

[Zbrafish] Low efficacy of splice-blocking morpholino - Zebrafish Danio Rerio Forum. Discuss zebrafish, aquarium culture and molecular biology lab specimens.


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Old 03-12-2007, 07:42 PM
Burdine, Rebecca D
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Default [Zbrafish] Low efficacy of splice-blocking morpholino




There may be maternal RNA in your embryos which your splice-blocking
morpholinos may not affect.

You don't say if you get a phenotype or not? If you do, perhaps you can
use both morpholinos together at lower doses to enhance the penetrance.

If you aren't getting a phenotype, you may not be able to induce one or
you may need to knockdown maternal message with an ATG or UTR
morpholino.

Becky

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-----Original Message-----
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Sent: Saturday, March 10, 2007 5:39 PM
To: [Only registered users see links. ]
Subject: [Zbrafish] Low efficacy of splice-blocking morpholino

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

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