APC-flox (Exon 1-14) 5'

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APC-flox (Exon 1-14) 3'

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APC-null

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Notes:

It is possible to get Apc-fle1-14 homozygotes and breed them, but the problem is that one can't distinguish heterozygous from homozygous conditional mutants by PCR genotyping. The WT band is always there in every genotyping protocol we've tried, or some contaminating band that is the same size as the wt band. We are not sure why. We tried lots of primer pairs and we do know that the targeting is correct, though, so no worries about that being the reason.

The way we breed homozygous mutants is by crossing to Apc-Min or other mutant Apc alleles. For example, progeny of Apc-fle1-14/Min x Apc-fle1-14/Min that didn't carry Min (Min genotyping protocol yields "WT") were Apc-fle1-14/fle1-14 homozygous.

Sorry that this sounds like a complete pain, and we seriously regret that we never got a genotyping protocol that worked. Find below files for sequence information for WT and Apc-fle1-14 allele in case you'd like to try to devise a genotyping protocol of your own. If you manage to find something that works, we'd love to hear about it. Good luck!

• APC_gen3prime_1lox1frt
• APC_gen3prime_wt
• APC_gen5prime_1lox
• APC_gen5prime_wt

10X PCR Buffer:

(store at -20ºC)