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Prejudice is observed on several levels throughout the course of the play. In the most obvious sense, the play deals with racial prejudice. While, conspicuously, the race of the accused is never certain, we do understand that he is a minority of some sort (in the 1957 film, the actor playing the accused was Italian), and this quickly becomes a heated issue among the jurors, especially for 9th Juror, who refers to the accused as "one of them."
Looking at prejudice in a larger sense, we find that, while maybe not racially driven, many of the jurors enter the jury room with preconceived notions and irrational ideas. 3rd Juror seems to be prejudiced against the accused simply because of his age, which seems to remind him of his estranged son. An interesting example of "reverse prejudice" is 8th Juror, who is initially sympathetic to the accused, not because of the evidence, but because he pitied his poor and troubled upbringing.