In the paper, it says these mice are from Jackson Labs. I've ordered mice from there before, and they usually have a "genotyping" tab that lists primers and all info to run a PCR. I've never used these mice but I've done this with others from Jax.

Thanks, Elyse. The genotyping protocol from JAX is a qPCR-based assay so I was looking for a std. PCR assay. I think I've found one though. Thanks for your help.

Did you get a working protocol? I have one that works with a standard PCR and gel electro that I am happy to share.

This is what we used.... Hope it helps.  We used amplitaq Gold MM.

Does anyone have a good protocol for genotyping APC flox mice from Jax (https://www.jax.org/strain/009045) C57BL/6-Apctm1Tyj/J? I am also looking for a std. PCR assay since the genotyping protocol from JAX may be a qPCR-based assay and did not work in our hands. Thanks, 

we are using dream taq for this PCR,

primers

wt APC mouse

5` -GCCATCCCTTCACGTTAG – 3`

 common antisense

5` -TTCCACTTTGGCATAAGGC – 3`

APC min mouse

5` -TTCTGAGAAAGACAGAAGTTA – 3`

products sizes:

·         WT : wt APC mouse p + common antisense (619 bp)

·         APC :APC min mouse +common antisense(600 bp +331 bp)

PCR reaction:

1.    For each reaction prepare the following mix according to the number of samples+2 (one for positive control, one for negative control)

·         Mix:

·         5 µl DreamTaq

·         2.4 µl UPW   (if the band size is different-like in cre- add 3.2 µl UPW)

·         0.2 µl primer 1 (1:10)

·         0.2 µl primer 2 (1:10)

·         0.2 µl primer 3 (1:10)

2.    Add 2 µl DNA from each sample to PCR tube-make sure you see the sample in the tube.

3.    Add 8 µl mix

4.    Put in the PCR machine and set the program according to your reaction

PCR:

step1 95°C      3min 

step 2 95°C      30sec

step 3 55°C      30sec

step 4 72°C      45sec

steps2-4      *  34 Cycles

72°C      10 min

16°C         Forever

good luck

Sharon's information is pretty similar to what we did.

For our work I varied the concentration of the primers so that the APC min band was not overpowered by the Wild Type band since they both occur when the PCR contains APC Min positive DNA.  I think we initially had issues with the PCR which resolved with primer concentrations.  We ran ours on a gel which worked great and did not use qPCR or the kit that Jackson sells.  The qPCR might be an advantage as this reaction is VERY sensitive to contamination.  Bleach forceps used for adding animal tissue to the DNA extraction.  We had lots of carry over w/o that step.  We also had contamination in the lab.  I did find that ALL APC Min mice had polyps in Small Intestines and found finally a source that corroborated that indeed ALL APC Min mice will have SI polyps.  Of course one has to sacrifice the mouse to examine the small intestine.  Perhaps there is also an age issue.  Our mice were about 16 or 20 weeks old at the time of necropsy.  You can double check your genotyping if needed by visual inspection.