While dissecting eyes I decided to genotype after fixing mouse eyes in 4% PFA for 15 minutes then subsequently washing in PBS for 5 minutes three times. Can you still genotype tissue after fixing it in PFA>?
It depends on how well your transgene is expressed. I've tried genotyping after fixation and usually doesn't work so well. You should always keep the yolk sac from embryos for genotyping.
You can genotype even ffpe = formalin-fixed paraffin-embedded tissue. Molecular pathologists do that all the time.
You might have to adjust your DNA extraction protocoll. There are extra extraction kits available for fixed tissue. And DNA quality is usually lower than from fresh tissue. So short PCR products would be recommended. But generally, it is possible. Depends of course also on what genotyping technology you want to use.
No other tissue left from your mice? Fresh tissue is of course recommended....
April and Lars-Erik thank you very much for your help. We did try and genotype the tissue from the eye including the optic nerve and we were able to get the genotype about 50% of the samples. Again, thank you for your expertise.
- Badges
- Science topic
- Similar topics
- Statistics
- Survey