Is there difference in the pronunciation of "to" and "too" and "two" ?
"to" and "too" and "two"に、発音の違いがありますか？

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There is no difference.

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No, all three are pronounced the same, but, as I'm sure you know, all three words have a different meaning.

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The main difference you should focus on is that "to" always has a weak accent, whereas "two" and "too" have strong accents.  So yes, we can hear the difference but it's a matter of stress; not the actual pronunciation of to/two/too, which is the same.

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They are not pronounced quite the same. 
The preposition "to" is pronounced the same as the words "too" and "two" only when it is followed by a word beginning with a vowel. It is pronounced as [tu:]. 
Before a word beginning with a consonant it is pronounced [tә], whereas the "o" is neutral or "schwa" sound.

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トゥー、トゥー、トゥー

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