[Zbrafish] Dechorionation of zebrafish embryos at gastrula stage (5.3-10hpf)

Jane via zbrafish%40net.bio.net (by coxj from slu.edu)
Fri Feb 3 16:59:21 EST 2012


Hi Farheen, We use Pronase, not proteinase K. Pronase is actually a protease from Streptomyces griseus, (sigma P6911). We use it at 1 mg/ml  and observe it under the dissecting microscope until the chorion crumples, around 5 min or less. For young embryos, you need to use a glass dish, because they stick to plastic. Hope this helps.

Regards, Jane 


Jane Cox
Asst. Res. Professor
Dept. Pharm. Phys, Sci.
Saint Louis University
1402 Sth. Grand Blvd.
St. Louis, MO 63104
(314) 977-6445
coxj from slu.edu




On Feb 3, 2012, at 1:10 AM, farheen f wrote:

> Hello,
> 
> Can anyone suggest the right concentration and incubation time to
> dechorionate embryos in the gastrula stage (5-10hpf) using proteinase
> K?
> 
> The protocol given at zfin suggests :
> 
> Digest zebrafish embryos with Proteinase K (10 ug/ml in PBSTw) at RT
> for 1 to 15 min depending on the stage: 1 cell to high for 1 min; 30%
> epiboly to 10 somites for 2-3 min; 10-20 somites for 3-4 min 24-32h
> for 5-6 min; 40-50 h for 10-15 min.
>                        Yet i am unable to get a good survival rate.
> 
> Thanks in advance,
> Regards,
> Farheen
> 
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