YES!!!!!!!
One night stay will get you front of the line 2 days if you do it right...
We arrived at the Portofino at 8:00 am from our other hotel, and left our luggage at Conceirge, and front desk gave us a temporary room card. We left hotel and hopped on boat that took us to the parks,and were there before opening.
The last day we took our luggage out of the room at 8:00 am and checked out, but did'nt turn in our room keys, had front of the line that last day too.
We were able to ride whatever we wanted, over ande over again. Rode MIB 7 times in a row one day!!!
I don't stay on-site at WDW, but the hotels are sooo nice at Universal, they aree worth it to me, and I'm a cheap, cheap, cheap-o!