I have a fully cycled 2.5g tank and I am going to set up a 5g tank, if I put all the substrate and water in the 5g, am I going to go throught the cycle all over again? Or can I transfer the fish from the 2.5g right away?
It certainly helps, but youre 5 gallon will only have the capacity to hold 2.5 gallons of bacteria. You probably would experience a mini-cycle. Start reeeeeal slow, especially in a tank that small. One fish at the most, to start out with.
The size of a bacterial colony is not dictated by the tank size, but by the fish load it has developed to support.
So yes, the 5 gallon will be already cycled, but only for the fishload the 2.5 gallon was cycled for. BTW - you don't need to transfer the water, but you do need to transfer the filter media.
You don't say how the 2.5 was cycled - with fish or fishlessly.