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Almost right away mabye a couple days to be absolutely positive. Boys have what looks like an outie belly button that is actually thier manhood. Our females seemed to have longer tails at a very young age not sure if this happens in general or just in our case, but reguardless it's easy to differentiate By lookin for the belly button "weenis". That being said, you shouldn't actually be touching the babies or mother at all the first 2 weeks after birth, if you do you increase the risk of mom abandoning or canabilizing the litter exponentially.
 

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You can tell from birth, but the babies should not be disturbed (this means peeking into nest/touching) until 2 weeks.
 
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