Quest For Glory IV: Nicolai's wife Anna, upon realizing that she is undead and has been dead for a long time.
Fate/stay night: Sakura pulls one of these ("IYAAAAAAAAAAAAAAAAA" which is Japanese for no) after she accidently stabs Shirou with a mana-draining projectile, nearly killing him. She feels incredibly guilty about this for the rest of the route.
Bob Page lets one off in the New Dark Age ending of Deus Ex.
In Baldur's Gate, if you have Khalid and Jaheira in your party, Khalid will yell one of these if Jaheira is killed.
Khalid: Ja-Jaheira! No! Noooooooo!
In Star Trek Online, after the player assembles the rescue beacon for their ship, Hakeev attempts to order his troops to kill them... Only to be foiled by the player being transported at the exact moment they would be fusilladed, in mission "The Coliseum", he delivers the Big "No!".
Montblanc in Final Fantasy Tactics Advance had this moment after fulfilling the request involving a townsgirl wanting to confess her true feelings, who actually doesn't exist, and in turn were a whole Moogle clan out to defeat him waiting in Materiwood. Only in the English version, though. The Japanese equivalent to this Big "NO!" is "?????~~~!!" (reads "Hidoikupo!!") which means TERRIBLE, KUPOOOOOO! with an addition of a This Cannot Be! before that.
Auron in Final Fantasy X let howl one when he finds that the sacrifice of his friends, Braska and Jecht, was for absolutely nothing and tries to unleash his fury on Yunalesca. Just to get beat, literally, to death. A "remembrance" echo as soon he saw Tidus having the same reaction over it he remembered what happened that time.
Eliwood does this twice in Fire Emblem: The Blazing Sword, first when his father dies and later when he realizes the dragon he just killed was Ninian.
Damon Gant does it twice — and that's before he actually slips into a Motive Rant.
Actually parodied at one point when The Judge does the Big "NO!" — and then stops and wonders why, since he's not the one in trouble.
In the same case, Phoenix lets out a Big "NO!" if you present to the judge at one point a piece of cloth with Ema Skye's fingerprints on it too soon. This makes everyone believe Ema really did accidentally kill the victim in the incident two years ago, making the trial impossible to win and you're notified that you eventually lost.
Ema herself let's off one when it is implied that She inadvertantly killed the very person she was trying to help.
So does Edgeworth, when he finds out he has unintentionally used forged evidence.
In Trials and Tribulations, when Edgeworth, who immediately caught on to how terrible being a defense attorney really is, just got through breaking three of Larry Butz's phsyche-lockes, being miserable the entire time through. It seemed he was about to get to the bottom of what Butz was hiding when suddenly he's encountered by five more! Cue the Big No. At that point, he decided to just take Butz as a witness.
Another particularly amusing one is when Matt Engarde shouts something to the like of "No!!!GuiltygUilTYGUILTYguiLTYgUiLty!!!" complete with alternating caps while ripping his face off.
Godot does such a moment in Case 2 of Trials and Tribulations WHILE DRINKING COFFEE. (This was notably not a Spit Take; he did this while chugging the coffee as fast as he could.)
Also, he also does the same thing in Case 3.
Phoenix himself lets out a big NO when Dahlia, while in Maya Fey's body, announces that Maya is dead.
Stages in Elite Beat Agents and Osu! Tatakae! Ouendan are touched off with the rescuee screaming in anguish (with the Big "HEEAALLLLLP!"/"OOOUUU-EENNN-DAAAAAN!", respectively), prompting an arrival and a dramatic entrance from the titular Big Damn Heroes.
Marcus Fenix in Gears of War 3 when he realises that Dom Santiago is about to sacrifice himself in order to assist his team's escape from the pipeline. Cue the sounds of the battle fading away to reveal an instrumental of Gary Jules' "Mad World".
Marcus: "Dom! What the fuck are you doing?!"
Dom: "Pullin' the plug on 'em, Marcus! Jump, will you?! Do it!"
Marcus:*realises what Dom is about to do* "DOM!! NO!!"
Max Payne breaks into the Big No right at the end of the opening level when he finds out that he was too late to save his wife.
This was parodied, along with everything else concerning the first Max Payne, in the Show Within A Show "Dick Justice" in Max Payne 2.
When Fox gets killed in Star Fox 64. Peppy and Falco don't use the Big No to a huge effect like everyone else does, but Slippy lets out a normal Big No.
One of the bosses does it too.
Subverted in Star Fox Assault'' by Wolf in Mission 7, where Fox rides Wolf's Wolfen. When Fox fails to protect the Wolfen, Wolf utters a soft, short "Curses." as his ship explodes, while Fox falls down screaming.
Snake shouts a Big "NO!" in Super Smash Bros Brawl if he's knocked out of the stage as a twinkle in the sky.
Peach goes "Noooo!" when she turns into a star too.
As does Sonic. In both the Japanese and English versions.
In Sonic Rivals and its sequel, Silver, Rouge, and shadow yell out a big "NO" when they take time to respawn.
In Dynasty Warriors, Sun Ce can die in Da Qiao's DW4XL stage. Right after he dies in her arms, she throws her head back and gives a really big NOOOOOOOOOOOOOOOOOOOOOOO!, complete with 360 camera action above her. Cue Roaring Rampage of Revenge. *
This is only in the Japanese version, though. The English version messed it up; even though the 360 camera action exists, the voice actress only gives a short "No...!"
Several variants of this trope in Dynasty Warriors 6, most in response to the same character's death. Your choice of player character determines exactly who is around to witness said death and shout their particular version of the Big No.
No slow motion but a really big NOOOOOO! can be seen from Garrett in Thief 2 (of Thief series) when Viktoria sacrifices herself in Soul Forge.
In the final episode of Season One, Sam lets out a normal Big No when Hugh Bliss separates Max's bliss and renders him kind and passive (the horror!), not to mention totally useless in helping to thwart Hugh's brainwashing plot.
Sam has two more in Season Three. The first is when Max's brain is stolen, and the second is just before Max transforms into an Eldritch Abomination.
In Heavenly Sword, Nariko yells this as Flying Fox throws Kai down with a noose around her neck, apparently killing her instantly. Although as it turns out Kai wasn't dead and eventually recovered.
Near the end of Myst III: Exile, villain/Woobie Saavedro does a less drawn-out version of this trope when you trap him behind the ice shields on Narayan. Repeatedly. In a dramatic, tormented, heart-wrenching manner. He does it again when you go to retrieve the MacGuffin from him, only this time, it's even more of a tearjerker.
Half-Life 2: Alyx has a big one at the end of Episode Two when she sees her father horribly killed in front of her.
Francis has one in Super Paper Mario after Peach (almost literally) nukes his laptop: "NOOOOOO! MY GRAPHICS CAAAAARD!"
Some of the characters in the Mario Kart series let out a Big No if they fall off the track.
Averted in Tomb Raider Underworld, when Lara's Doppleganger shoots Alister, Lara goes over to him, when he actually dies, Lara just utters a sharp 'Dammit!', then carries his body out of Croft Manor, now on fire.
Sora's going to do his Heroic Sacrifice. And everyone around him goes "NOOOOOOOOOOO! SOOOOOOOOOORA!!!"
In Chain of Memories: Reverse Rebirth, the Riku Replica gives one it's decided to have NaminÚ give him a false set of memories and make him forget that he's not the real Riku.
Demyx gives one of these when you beat him. "Nooooo waaaaayyyy!"
The good ending of Disgaea 3: After the final battle, Mao puts up with Aurum's speech before pulling out his sword and preparing to behead the Super Hero... before deciding that he'd be better served as a research specimen. Aurum is naturally distraught at this last-minute rejection of the death he always wanted.
Mao:(evil laugh) Have you forgotten? I am a demon! Your logic won't work on me! TO MY LAB! (guess what Aurum says next)
Video games in general often use either this trope or Say My Name in place of the player-character's death scream. This is especially apparent in Sonic the Hedgehog games.
For example, in Sonic the Hedgehog (2006), Sonic and Shadow both yell "No!" (the latter much more dramatically) upon dying, while Silver cries "No... not now..."
In the same game, he also lets off a half-big No in the opening when Merlina warps them away before Sonic gets the chance to recover his second Chili Dog, however the subtitle is more shaped to that of a Big "NO!"
Not to mention Elise's literally explosive Big No when Sonic is killed.
In Sonic Adventure 2, in the Dark Side story, Sonic busted the Restraining Bolt on the Egg Golem boss. After throwing a small fit, the Golem turned to Eggman and attacked him in dramatic slow-mo, accompanied by Eggman shouting in a deep voice, "NOOOO...!!!"
In Sonic Rush Adventure, when your craft takes on damage, Tails says "NO!"
Used during several points in Gabriel Knight: Sins of the Fathers, when Gabriel first meets Tetelo in the Bayou, when Wolfgang rips out his own heart so Gabriel can get the talisman, and after Malia sacrifices herself to finish Tetelo.
One of the achievements in Team Fortress 2 is titled "Dr. Nooooo". To accomplish it, you have to backstab a Medic who's about to deploy an Ubercharge; as that takes time to build up and would have rendered the Medic invulnerable if he'd pulled it off, they probably would let out a Big No at their monitor.
One the Spy's Bond One Liners after being ▄bercharged by a Medic is "They should call you whiners 'Dr. Noooooooooooo'!"
Interesting variation in Kings Quest VI Heir Today Gone Tomorrow: "That's odd. Where did she..." (The floor drops out from under Alexander's feet.) "...GOOOOOOOOOO?!?!" Played straight later on by Cassima, who gives an anguished "Alexander, noooooooooooo!" if Alexander is killed by Shamir Shamazel in the endgame confrontation.
In the second episode of the Fan SequelThe Silver Lining, Alexander lets out a Big "NO!" just before being energy zapped by Manannan in a vision Graham experiences.
The Commodore 64 port of 1984's Impossible Mission rewards the player who completed the game by revealing the villain, accompanied by a digitized, plaintive "No. No. NO! The ZX Spectrum version (when it allowed the player to reach the ending, without tripping over the Game-Breaking Bug) has the picture of the villain screaming "NOOOOOOOOO!" (in the image), accompanied by a shaking screen, white noise and stripy border. You can go to Speccy Spoilers and grab a copy of this ending to see it for yourself (for an emulator on which to run that file, just visit the base of that URL)...
The original Neverwinter Nights has Fenthick utter a beautiful exemplar at the end of the first chapter: "Desther! NOOOOOOOOOOO!"
The girls of Persona 4 are known for their terrible cooking skills; the boys called anything they cook Mystery Food X ever since their first taste. The boys were terrified to learn that the girls baked a Christmas cake to celebrate little Nanako recovering after getting sick from being kidnapped into the Midnight Channel. Kanji lets out a NOOOOOOO! when she tries the cake before they could. The cake was delicious.
You also encounter one in text form upon reaching a certain floor of the Void Quest dungeon.
Tatsuya does it at the end of Innocent Sin/in the flashback of Eternal Punishment (although it's never seen in Innocent Sin, it's shown in the flashback) in Persona 2. It sort of turns a really emotional scene into a big Narm moment due to the bad English voice acting.
Plants vs. Zombies uses this in the Game Over screen, which says "The Zombies Ate Your Brains".
There is also a black dragon named Nihil the Banished that may spawn randomly during a quest involving warping nether dragon eggs. When he spawns right before the player, he will say something about world domination, but if the player uses the device again on him, he will be banished again while yelling NOOOOOooooo....
"NOOOO!!! A curse upon you, interlopers! The armies of the Lich King will hunt you down. You will not escape your fate..."
Interestingly enough, the second-to-last boss in the Nexus yells "Nooo!" when he's engaged, rather than when he's killed.
Because it's basically a requisite for Star Wars, The Force Unleashed has Starkiller let one out during his Heroic Sacrifice in the good ending. He also does it twice in the bad ending (one when he sees all his friends are dead and one when he gets crushed by a space ship).
Before that, Kota lets one out when he realizes that Palpatine is trying to turn Starkiller to the Dark Side.
In The Force Unleashed II, Starkiller lets one out when he fails to stop Boba Fett's ship, which has Juno Eclipse captive, from taking off and escaping.
In Zork: Grand Inquisitor, when the player puts a Zorkmid in a vending machine, the snacks inside inch forward but don't come out, prompting a Big No from Dalboz (though he's a lantern now, so even if they did come out he couldn't actually eat them).
In Dark Forces, General Rom Mohc gives a really good one upon his defeat.
If you manage to get a mod that changes all zombies in Left 4 Dead 2 to Jimmy Gibbs Jr, or manage to find a way to spawn him, Ellis shouts NNNNOOOOOOOO! when he sees his hero zombified. Its funnier that the other survivors don't care, like Coach saying sorry about the car and Rochelle saying Dibs on Gibbs or HOLY SHIT whats that.
Also, any time a Smoker grabs you in the first Left 4 Dead.
Homeworld: Cataclysm contains a particularly entertaining variation at the very end of the last mission, when the Naggarok, big bad and 'parent' of the game's nightmare fuel main enemy (aptly named the Beast), is destroyed. The "Nooooooooooo!" goes on for a good long while, even after the boss itself has been reduced to cosmic dust.
In Master Of Orion II: Battle at Antares, you hear this when a ship bearing a leader is killed in battle, and the dead ship's owner hasn't researched Survival Pods tech, which allows you to retrieve a leader as long as at least one ship survives the battle.
Kilrathi pilots in Wing Commander II sometimes do this when they get shot down. So do some of the hostile human pilots in Wing Commander IV.
In the Quake series whenever you fall into an instant death pit.
Armed Police Batrider's announcer shouts "NOOO!" if you get a Game Over or miss a 10,000-point medal.
In The Godfather game, Monk Malone yells one after a string of smaller "no"s when Frankie Malone dies after being abducted by Tattaglia goons, even hurling a church pew by himself in his rage and sorrow.
Happens fairly often in Supreme Commander when you kill an enemy commander. The most memorable is certainly Marxon's at the end of the Aeon campaign.
Spyro the Dragon has one of these in A New Beginning, when Sparx is about to be stepped on ("NOOOOOO—AAAAHHHHHHHH!", he was breathing fire for the first time), and in Dawn of the Dragonwhen Ignitus dies.
Dark Messiah of Might and Magic includes a pretty corny one in the most illogical of the game's possible endings. If you cleanse yourself of Xana's demonic possession, then go ahead and decide to free Daddy Archdemon at the end of the game, Leanna responds to this with a big no.
In Chrono Cross, Radius screams an all-caps Big "NO!" after the evil power of the Masamune causes him to kill his best friend Garai.
Ace Dixon gets a few of these in Crimson Skies. The second and fourth times you shoot him down, you'll be thanked over the radio with this:
In Luminous Arc 2, Rina gave a big one after Steiner died in her arms.
In SNK Gals Fighters, this is one of Yuki's moves. Very useful for offense and defense.
Sieg from Suikoden Tierkreis gives this on a few occasions, but a really big one comes out right after Dirk's death.
In the endgame of Monkey Island 2: LeChuck's Revenge, Guybrush and LeChuck do a nearly word-for-word recital of the classic scene from The Empire Strikes Back, which, naturally, includes the Big "NO!" It's every better in the Special Edition version, where Dominic Armato (as Guybrush) performs his lines exactly the same way as Mark Hamill did in Empire, up to and including the Big "NO!"
A lot of Artix Entertainment villains tend to use these often when they're defeated, like Ledgermayne when it's being converted back into the raw mana from which it was made by the Supreme Arcane Staff and Kitsune when he ends up being sucked into and sealed away in a rift leading him back to the Yokai world, that while saying a variation of "This Cannot Be!" in the process.
Kitsune: (as he is about to run into the rift) No! Not like this! It can't end like this! I won't go back to the Yokai world! NOOOOoooooooooooo! (gets sucked into the rift)
Even Dr. Voltabolt in the Adventure Quest Worlds Mogloween events has one as well after his Dental Driller is destroyed.
Dr. Voltabolt: Nooo! My beautiful machines! You haf ruined my planz! Hero: Serves you right! You were trying to ruin Mogloween for everyone!
Blizzy lets out one after Lionfang performs his Kick the Dog moment of burning the gifts, and another one after he shatters the snow globe containing the Spirit of Frostval and lays bare his intent to make the world suffer for "surrendering to evil" by destroying Frostval (which, fortunately, can't happen so easily, as the Spirit returns to Lionfang's shock and disbelief to set things right again).
In the finale of the Doomwood saga, Vordred uses his dreaded Voiduminance Necrot-Morph spell, the same spell that he used to turn the Paladins of Doomwood undead, on Artix. Cue the Big "NO!" from the Hero. Fortunately, the spell has no effect on Artix — but that's because Artix is the Champion of Darkness.
Portal 2: During the core transfer, GLaDOS yells probably the most epic autotuned NOOOOO ever.
Dragon Age II: In Act 3, when Anders blows up the chantry, Sebastian lets out a narmy "MAKER NOOOO!"
Putt-Putt Travels Through Time: After Pep gets sucked into the time machine and gets lost: "Pep! NOOOOOOO!"
In the first Pajama Sam game, Otto lets one out as he goes down a waterfall.
Double Switch: Eddie yells this a few times when you trap him at the end of the game.
Klogg from The Neverhood utters a Big No TWICE during the good ending. First, he yells it when Klaymen is about to put the crown back on Hoborg's head:
This can happen five times in Saints Row The Third: four times during one of the possible final missions courtesy of the Boss, consecutively as all of his/her buddies die, and once said by Killbane if you decide to unmask him.
Already having been tortured and beaten senseless after his capture, this is McNeill's reaction in the final Nod cutscene of Command And Conquer Tiberian Sun when Slavik shows him the destruction of the Philadelphia on the viewscreen.
Age Of Mythology has the [[Big Bad]] Gargarensis who has this as a stock phrase when you foil one of his plans.
Happens in OddworldMunch's Oddysee when glukkon Lulu realizes that, due to forces behind his understanding (EI, Abe's possession), he's spent his entire fortune on a can of gabiar, which is basically caviar IN SPACE. His anguished cries can be heard across the countryside ("What was that?" "Somebody's really pissed...") and is even written down in the newspaper article.
Viewtiful Joe: Captain Blue delivers one of these, when he is defeated in his King Blue form.
In I Miss The Sunrise, this is Ros' reaction to witnessing the destruction of the Inquiry.
Not a video game exactly, but closely related. In the Pinball game Monster Bash, the Bride of Frankenstein does one of these when "Ball and Chain" mode is activated.
In "Cirqus Voltaire" (sic), the Ringmaster sometimes gives one when he's defeated.
The genie will also sometimes give one in Tales of the Arabian Nights when he's defeated in "Fireball" mode.
Asura of Asura's Wrath screams this in Episode 12 as Olga and her fleet are launching the bombing that kills the girl who reminds him so much of his daughter. Considering he has no arms to counter the bombs with, there's not much else he can do at this point.
Brain Dead 13: Dr. Neurosis does this on noticing that Fritz has been tricked into completely damaging his computer and machinery after Lance defeats him; this is followed by the Big Bad's shout of "Fritz, you imbecile!"
In the climax of your first dragon battle in The Elder Scrolls V Skyrim, Mirmulnir, the dragon you fight, lets out a great big NIIIIIIIID! upon realizing too late that you are the Dragonborn (a mortal with the soul of a dragon, capable of taking the soul and power of any dragon he or she slays) when you deliver the final blow to him.
In Xenosaga, Albedo, the Ax CrazyBig Bad, does one in the second game when he finds out that he, being virtually immortal, will eventually outlive his brothers...
In Cardinal Sins: Judgement Silversword Recycle Edition, one mission tasks you with collecting One Ups. In this particular stage along, if you shoot a 1-up too many times, it will explode, and a "NO!" message will pop up.
Played for laughs in Puyo Pop when Arle yelled one after being told that her skeletal first opponent had made the tea he'd served her with used teabags - then commented that it was "not that bad."
In Werewolf: The Lost Warrior, shouting "NOO!" at your enemies is the strongest attack you can use in human form.