I kinda agree with a few other posters on this thread that there can never truly be a definitive Spider-man story but I'll play along. LOL

A definite story, one that comes close for me is: Spider-Man: Life Story. I recently read the whole thing and I kinda loved it. Spider-Man's whole story from the 60s all the up till now. He got his powers, He did the superhero thing. He got OLD, He married, he had kids, and he inspired others and of course HE died heroically.

My all time favorite Spider-Man story: Amazing Spider-Man Vol 2: Happy Birthday! Issues #57, 58, and renumbered to 500.

I'm actually Amazed that not alot of ppl give this Arc the love that it deserves. It's my all time favorite Arc with 500 being my all time favorite issue. Dormammu has invaded Earth and instead of Waiting on Doctor Strange to handle it the Heroes of 616 fight the good fight and Reed Comes up with some science mumbo jumbo that actually destroys everything LOL. But Strange was able to reach to Spidey, and fling him into the past or something like that. And so he has to fight every single villain, relive every single moment of heroism and tragedy in his life in order to 'catch' back up to the moment Reed destroyed everything.

It's an awesome story plus this story is where the Spider-Man the last stand suit originated.