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If that doesn't help, use the pause menu tuner to ensure you really are tuned to the correct note as the tuner before songs is very loose with its accuracy. During a song press Esc and you'll see the chromatic tuner in the lower left. From the main menu, I believe you can see the tuner in the Tools screen.
If your open string is then recognized but notes higher up the fretboard are not, you might have a problem with intonation, which I've learned is a big issue for cheap equipment. You can try to correct it yourself after watching any of the number of instructional videos on YouTube.
Also, there's this guide if you want to ensure a good setup all around. http://forums.ubi.com/showthread.php/802854-Rocksmith-2014-PC-Configuration-and-FAQ-Forums
I have a similar problem tuning my guitar. All worked well for weeks until now. I had to do the same work around as GrumbleMutt mentioned above (initially to get the sound working), but recently it all went bad. On a Drop D tuning song,It asks me to tune the "D" string, then it jumps to quickly to the "e" string, strum , it then says you guitar is tuned. Its not even close. I believe its a software issue.
Can you reload Rosksmith 2014 in Steam, if so, How?