Revealing mistake: While Indy and Marion are in the Well of Souls, and they encounter the snakes, Indy falls to the ground only to be confronted by a hissing Cobra rearing its head. Look carefully and you'll see the snake's reflection on the safety glass between it and Indy. Briefly you can also see the torch's reflection while he's waving it around. [This has been corrected in the new DVD set. You can only see it if you have the original VHS. I think it's visible in the "making of" on the DVD as well.]
Continuity mistake: When Indy goes to save Marion in the tent he removes the handkerchief gag. But when he decides to leave her there, he puts the gag back on and some of her hair gets caught underneath the gag on one close up shot from the left. But when the camera goes to the back of Indy, her hair is no longer caught underneath the gag.
Visible crew/equipment: After Marion supposedly dies in the truck explosion and Indy is drinking outside at the bar, watch to the left of him. You can see a man wearing a modern t-shirt and jeans walking through the crowd.
Visible crew/equipment: As soon as Marion gets locked in the cockpit of the Nazi plane, you can see a crew, a blue van, a ladder, and a man with a white sleeveless shirt and a big brimmed hat all in the reflection of the glass.
Continuity mistake: Near the beginning of the movie where Indiana is running away from the tribe of South American Indians after Belloq robs Indiana of the gold statue, in one shot you can see that the Indians are only a few meters behind Indiana, but in the next shot, they are at least 50 meters behind.
Continuity mistake: In the Himalayan bar scene when the sherpas leave, Marion starts to pick up the shot glasses and puts them on a tray - there's a large group left at one end of the table, one in the middle, and two by a bowl at the other end. When she returns for the rest, the two by the bowl have vanished.
Continuity mistake: When the village crowd separates and reveals a standoff between Indy and the black-robed swordsman, the rear shot of Indy shows his hip-bag to the rear of him, the frontal shot of Indy shows his hip-bag on the forward part of his hip.
Revealing mistake: When Indy fights the driver of the Nazi truck, he gets thrown through the windshield, and eventually ends up underneath the truck. When he is climbing underneath the truck, heading towards the back of it, you can see the trench that was dug down the middle of the road to provide the stuntman (this being one of the few stunts that Ford didn't do himself) with the extra room needed to do this feat. (This is backed up by DVD commentary. The director stated that the truck was too low to the ground so they had to dig the trench in order for there to be enough room for the stuntman).
Revealing mistake: When Indy jumps across the pit to pursue the spy that took the idol, he barely makes it across, grabbing the ledge on the opposite side with his hands. When he pulls himself up, he kicks loose a rock and the entire "solid" rock wall shakes like it's nothing more than a foam wall.
Factual error: At the beginning, while Indy and his two native guides are slogging through the Peruvian rain forest, one of the birdcalls in the background is that of an Australian kookaburra.
Revealing mistake: After Indy gets behind the wheel of the truck containing the ark, he jerks the steering wheel to the right to run the pursuing jeep off the road. He then jerks the wheel to the left to hit an upcoming motorcycle. In neither case do you see his body sway with the presumably drastic swerve of the truck. Apparently this was filmed in a stationary vehicle, or one in tow that was fitted with a prop steering wheel.
Continuity mistake: When Belloq is talking to Indy and Marion in the Well of Souls, he's shown holding his hat with his right hand. But in the only over-the-shoulder shot (looking into the Well of Souls) the hat is laying on the ground and Belloq tosses a stone down with his right hand. Then the hat is back.
Revealing mistake: Watch the shadow of the stone Indy pushes out to escape from the Well Of Souls - it bounces down the sand dune, exactly the way a large rock shouldn't.
Factual error: It should be a tricky task to put an object the size of the Ark on board a German VIIC submarine; perhaps you can find enough place inside but how do you manage to introduce that bulky case through the narrow hatches of the sub? Even the torpedo load hatches were just a little more than 21 inches in diameter.
Continuity mistake: At the beginning of the film, Indiana Jones uses his whip to disarm a man with a handgun. The scene showing this action includes several quick shots, but as a result of the use of these multiple shots it can be seen that the whip descends twice. (Slow motion helps, but isn't required.)
Factual error: Indy's satchel is a Mk VII gas mask bag, which was created for use in World War II, so it didn't even exist at the time the film takes place (1936). The similar Mk V existed at the time, though.
Factual error: When Indiana Jones is fighting the big German by the airplane, the German punches Indiana Jones on his right cheek. Indy's face and body are both knocked down to the right, in the direction of the punch. When you watch this, you can see it's impossible to be knocked down in such a way.