ASCII reserves the first 32 codes (numbers 0–31 decimal) for control characters: codes originally intended not to carry printable information, but rather to control devices (such as printers) that make use of ASCII, or to provide meta-information about data streams such as those stored on magnetic tape. For example, character 10 represents the "line feed" function (which causes a printer to advance its paper), and character 8 represents "backspace". RFC 2822 refers to control characters that do not include carriage return, line feed or white space as non-whitespace control characters.^[30] Except for the control characters that prescribe elementary line-oriented formatting, ASCII does not define any mechanism for describing the structure or appearance of text within a document. Other schemes, such as markup languages, address page and document layout and formatting.

The original ASCII standard used only short descriptive phrases for each control character. The ambiguity this left was sometimes intentional (where a character would be used slightly differently on a terminal link than on a data stream) and sometimes more accidental (such as what "delete" means).

Probably the most influential single device on the interpretation of these characters was the ASR-33 Teletype series, which was a printing terminal with an available paper tape reader/punch option. Paper tape was a very popular medium for long-term program storage up through the 1980s, lower cost and in some ways less fragile than magnetic tape. In particular, the Teletype 33 machine assignments for codes 17 (Control-Q, DC1, also known as XON), 19 (Control-S, DC3, also known as XOFF), and 127 (DELete) became de facto standards. Because the keytop for the O key also showed a left-arrow symbol (from ASCII-1963, which had this character instead of underscore), a noncompliant use of code 15 (Control-O, Shift In) interpreted as "delete previous character" was also adopted by many early timesharing systems but eventually faded out.

The use of Control-S (XOFF, an abbreviation for transmit off) as a handshaking signal warning a sender to stop transmission because of impending overflow, and Control-Q (XON, "transmit on") to resume sending, persists to this day in many systems as a manual output control technique. On some systems Control-S retains its meaning but Control-Q is replaced by a second Control-S to resume output.

Code 127 is officially named "delete" but the Teletype label was "rubout". Since the original standard gave no detailed interpretation for most control codes, interpretations of this code varied. The original Teletype meaning, and the intent of the standard, was to make it an ignored character, the same as NUL (all zeroes). This was specifically useful for paper tape, because punching the all-ones bit pattern on top of an existing mark would obliterate it. Tapes designed to be "hand edited" could even be produced with spaces of extra NULs (blank tape) so that a block of characters could be "rubbed out" and then replacements put into the empty space.

As video terminals began to replace printing ones, the value of the "rubout" character was lost. DEC systems, for example, interpreted "Delete" to mean "remove the character before the cursor," and this interpretation also became common in Unix systems. Most other systems used "Backspace" for that meaning and used "Delete" to mean "remove the character at the cursor". That latter interpretation is the most common today.

Many more of the control codes have taken on meanings quite different from their original ones. The "escape" character (code 27), for example, was originally intended to allow sending other control characters as literals instead of invoking their meaning. This is the same meaning of "escape" encountered in URL encodings, C language strings, and other systems where certain characters have a reserved meaning. Over time this meaning has been co-opted and has eventually drifted. In modern use, an ESC sent to the terminal usually indicates the start of a command sequence, usually in the form of a so-called "ANSI escape code" (or, more properly, a "Control Sequence Introducer") beginning with ESC followed by a "[" (left-bracket) character. An ESC sent from the terminal is most often used as an out-of-band character used to terminate an operation, as in the TECO and vi text editors.

The inherent ambiguity of many control characters, combined with their historical usage, created problems when transferring "plain text" files between systems. The clearest example of this is the newline problem on various operating systems. On printing terminals there is no question that you terminate a line of text with both "Carriage Return" and "Linefeed". The first returns the printing carriage to the beginning of the line and the second advances to the next line without moving the carriage. However, requiring two characters to mark the end of a line introduced unnecessary complexity and questions as to how to interpret each character when encountered alone. To simplify matters, plain text files on Unix and Amiga systems use line feeds alone to separate lines. Similarly, older Macintosh systems, among others, use only carriage returns in plain text files. Various DEC operating systems used both characters to mark the end of a line, perhaps for compatibility with teletypes. This de facto standard was copied into CP/M and then into MS-DOS and eventually into Microsoft Windows. Transmission of text over the Internet, for protocols as E-mail and the World Wide Web, uses both characters.

The DEC operating systems, along with CP/M, tracked file length only in units of disk blocks and used Control-Z (SUB) to mark the end of the actual text in the file (also done for CP/M compatibility in some cases in MS-DOS, though MS-DOS has always recorded exact file-lengths). Text strings ending with the null character are known as ASCIZ or C strings.