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Isolation of skin from live skin-specific knockout mouse


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#1 scienceisfun

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Posted 19 October 2009 - 05:34 PM

Hi All,

Does anyone have advice on how I might isolate a skin sample for genotyping a skin-specific knockout mouse in a non-invasive manner (these mice are not able to be culled)? We will be performing skin carcinogenesis studies on these mice and I would like to avoid having to punch biopsy them for genotyping. Does anyone know an isolation protocol from the tail that is highly specific for skin?

Thanks in advance

#2 Rsm

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Posted 02 November 2009 - 09:29 PM

I'd suggest you to use the ear for genotyping. Tail is rather difficult to digest... Just cut a piece of ear when you tag them.
Cheers
Minna
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#3 scienceisfun

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Posted 02 November 2009 - 10:00 PM

I'd suggest you to use the ear for genotyping. Tail is rather difficult to digest... Just cut a piece of ear when you tag them.
Cheers
Minna


Hi Minna,

Thanks for your reply. The difficulty is not in tissue type (we genotype from both ear notch and tail successfully, tail tip is actually easier than ear notch) but in tissue specificity. The ear has other tissue present that confuses the genotype. We need a pure keratinocyte sample to perform the genotype.

#4 bob1

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Posted 03 November 2009 - 03:39 PM

The only method I can think of would be to do a partial digest of the biopsy and label for keratinocyte markers then FACS... rather laborious but it should work.




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