There are other federal trial courts that have nationwide jurisdiction over certain types of cases, but the district court also has jurisdiction over most of those types of cases, and the district court is the only one with jurisdiction over criminal cases and the only one where a trial can be to a jury instead of just a judge. The Court of International Trade addresses cases involving international trade and customs issues. The United States Court of Federal Claims has jurisdiction over most claims for money damages against the United States, including disputes over federal contracts, unlawful takings of private property by the federal government, and suits for injury on federal property or by a federal employee. The United States Tax Court has jurisdiction over cases for a refund of taxes.

The number of judges in each district court (and the structure of the judicial system generally) is set by Congress, and the Senate has to approve each appointment of someone to be a judge; the President appoints all judges, so they virtually always belong to the same political party as the President. Judges are appointed for life, but a judge who has reached the age of 65 (or has become disabled) may retire or elect to go on "senior status" and keep working. Such "senior" judges are not counted in the quota of active judges for the district and do only whatever work they are assigned by the chief judge of the district, but they keep their offices (called "chambers") and staff, and many of them work full-time. A federal judge is addressed in writing as "The Honorable Jane Doe" or "Hon. Jane Doe" and in speech as "Judge" or "Judge Doe" or, in a courtroom, "Your Honor."

To file a civil case (that is, "sue someone") in federal district court, a person must have a reason why a federal court, instead of a state court, should adjudicate the dispute. By law, the four bases for federal jurisdiction (the power to hear and decide a case) are: United States as a plaintiff; United States as a defendant; "federal question," which means the complaint is based on a federal law (which may be the Constitution or a statute); and "diversity of citizenship," which means the plaintiff, or person suing, and defendant (person being sued) live in different states and the "amount in controversy" is more than the statutory minimum, which is currently $75,000. Thus, not every legal dispute can be litigated in federal court, hence the expression "make a federal case out of it."

Although in matters of civil law there are often parallel federal and state laws, providing an aggrieved party with a choice of venue, there are some matters which may only be adjudicated in the Federal courts; these include most intellectual property questions and matters related to international relations. In some situations, Federal law provides both for the exclusive jurisdiction of Federal courts and for the immunity of the defendant from the power of those courts. One example of this is patent-infringement claims against a state government: only the Federal courts may hear patent cases, but the states have sovereign immunity from such suits. Although a state may choose to waive its immunity in such a case and allow it to proceed to trial, if it does not do so, the plaintiff has no recourse. (This doctrine was reaffirmed in a pair of Supreme Court cases titled Florida Prepaid Postsecondary Education Expense Board v. College Savings Bank.)

A formal ruling by a district court in either a civil or a criminal case can be appealed to the United States court of appeals in the federal judicial circuit that court is in.

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