"Azumanga Web Daioh" is a comedic slice-of-life anime that centers on the daily lives of a group of high school girls and their interactions with each other, their teachers, and the world around them. The narrative unfolds in a loosely episodic format, capturing the mundane yet humorous moments of their school years. The main characters include Chiyo Mihama, a child prodigy who skipped several grades and is now attending high school at the age of 10; Osaka, a transfer student from Osaka whose slow, dreamy demeanor often leads to surreal and absurd conversations; Tomo Takino, a hyperactive and competitive girl who frequently clashes with others; Koyomi Mizuhara, a studious and level-headed class representative; Ayumu Kasuga, nicknamed "Osaka" due to her similar personality to the transfer student; and Sakaki, a tall, quiet girl with a love for cats, though her intimidating appearance often leads to misunderstandings.

The series also features their eccentric teachers, particularly Yukari Tanizaki, their irresponsible and often immature homeroom teacher, and Minamo Kurosawa, a more composed and responsible physical education teacher who frequently clashes with Yukari. The interactions between the students and teachers, as well as among the students themselves, form the core of the narrative. The humor arises from the characters' distinct personalities and their reactions to everyday situations, ranging from school festivals and exams to summer vacations and cultural events. The anime does not follow a strict overarching plot but instead focuses on vignettes that highlight the quirks and dynamics of the group. The tone is lighthearted, with occasional moments of surreal comedy and heartfelt camaraderie, making it a quintessential example of the slice-of-life genre.