Duke Nukem has 2 main multiplayer modes such as Co-op and deathmatch, each mode can have up to 8 players working together or killing each other in a free for all match up. This is available for the PC version and the Xbox Live version as well. There are leaderboards on the Xbox 360 version which show what rank you are and how many kills you have scored on each mode. All the levels as seen in the story can be played on multiplayer which is 4 Episodes worth of levels unless you have the PC version in which you can add more to the list of levels available to play.

Dukebots can be used in multiplayer games by typing in the command line "Duke3d /l#(level number) /v#(episode number) /a(Use fake multiplayer bots) /q#(number of dukebots to play with). They exist on the Atomic Edition, Duke Nukem 64, and Duke Nukem Forever. They are utilized for deathmatch games. The difficulty of the bots cannot be changed in Duke Nukem 3D Atomic Edition. They are not scripted correctly for a variety of reasons: being able to always hit their opponents with whatever weapon they have, even over extremely long distances, with every weapon they use, strafe very well, move very fast, fire very fast, occasionally get stuck in some places, killing themselves (Using explosives, being crushed, etc), always knowing where their adversaries are hiding, are overkill with high powered weaponry (Shotgun, Chaingun, RPG, Microwave Expander, and Devastators), and they do not know how to place Laser Tripbombs, since these were bots used to test how deahmatches were fought. They are noticably seen moving erraticaly when a player "spies" them (through a security camera, staying out of sight from them). They always stay at an end of a sector that a nearby player is in, until a player nearby moves to a different sector, causing the dukebot to come after them. They do not perform very good in Cooperative as they choose to kill their teammates instead of the aliens as their purpose for deathmatch was to target players.

In Duke Nukem 64, the difficulty of the bots can be changed to make the bot weaker or stronger. On lower difficulty levels, the bots will fire slowly and are less accurate, but they still keep the ability to always know where players are. On higher difficulty levels, they will be very smart, fire much faster, and are more accurate.

Some modifications feature dukebots that have scripts which work correctly, like in Duke Nukem Arena.

And you can play Duke3d online in Meldown and Dukematcher ,