The mimetypes module in the standard library will determine/guess the MIME type from a file extension.

If users are uploading files the HTTP post will contain the MIME type of the file alongside the data. For example, Django makes this data available as an attribute of the UploadedFile object.

More reliable way than to use the mimetypes library would be to use the python-magic package.

import magic
m = magic.open(magic.MAGIC_MIME)
m.load()
m.file("/tmp/document.pdf")

This would be equivalent to using file(1).

On Django one could also make sure that the MIME type matches that of UploadedFile.content_type.

13 year later...
Most of the answers on this page for python 3 were either outdated or incomplete.
To get the mime type of a file I use:

import mimetypes

mt = mimetypes.guess_type("https://www.w3.org/WAI/ER/tests/xhtml/testfiles/resources/pdf/dummy.pdf")
if mt:
    print("Mime Type:", mt[0])
else:
    print("Cannot determine Mime Type")

# Mime Type: application/pdf

Live Demo

From Python docs:

mimetypes.guess_type(url, strict=True)

Guess the type of a file based on its filename, path or URL, given by url. URL can be a string or a path-like object.

The return value is a tuple (type, encoding) where type is None if the type can’t be guessed (missing or unknown suffix) or a string of the form 'type/subtype', usable for a MIME content-type header.

encoding is None for no encoding or the name of the program used to encode (e.g. compress or gzip). The encoding is suitable for use as a Content-Encoding header, not as a Content-Transfer-Encoding header. The mappings are table driven. Encoding suffixes are case sensitive; type suffixes are first tried case sensitively, then case insensitively.

The optional strict argument is a flag specifying whether the list of known MIME types is limited to only the official types registered with IANA. When strict is True (the default), only the IANA types are supported; when strict is False, some additional non-standard but commonly used MIME types are also recognized.

Changed in version 3.8: Added support for url being a path-like object.

Python bindings to libmagic

All the different answers on this topic are very confusing, so I’m hoping to give a bit more clarity with this overview of the different bindings of libmagic. Previously mammadori gave a short answer listing the available option.

libmagic

• module name: magic
• pypi: file-magic
• source: https://github.com/file/file/tree/master/python

When determining a files mime-type, the tool of choice is simply called file and its back-end is called libmagic. (See the Project home page.) The project is developed in a private cvs-repository, but there is a read-only git mirror on github.

Now this tool, which you will need if you want to use any of the libmagic bindings with python, already comes with its own python bindings called file-magic. There is not much dedicated documentation for them, but you can always have a look at the man page of the c-library: man libmagic. The basic usage is described in the readme file:

import magic

detected = magic.detect_from_filename('magic.py')
print 'Detected MIME type: {}'.format(detected.mime_type)
print 'Detected encoding: {}'.format(detected.encoding)
print 'Detected file type name: {}'.format(detected.name)

Apart from this, you can also use the library by creating a Magic object using magic.open(flags) as shown in the example file.

Both toivotuo and ewr2san use these file-magic bindings included in the file tool. They mistakenly assume, they are using the python-magic package. This seems to indicate, that if both file and python-magic are installed, the python module magic refers to the former one.

python-magic

• module name: magic
• pypi: python-magic
• source: https://github.com/ahupp/python-magic

This is the library that Simon Zimmermann talks about in his answer and which is also employed by Claude COULOMBE as well as Gringo Suave.

filemagic

• module name: magic
• pypi: filemagic
• source: https://github.com/aliles/filemagic

Note: This project was last updated in 2013!

Due to being based on the same c-api, this library has some similarity with file-magic included in libmagic. It is only mentioned by mammadori and no other answer employs it.

2017 Update

No need to go to github, it is on PyPi under a different name:

pip3 install --user python-magic
# or:
sudo apt install python3-magic  # Ubuntu distro package

The code can be simplified as well:

>>> import magic

>>> magic.from_file('/tmp/img_3304.jpg', mime=True)
'image/jpeg'

You didn't state what web server you were using, but Apache has a nice little module called Mime Magic which it uses to determine the type of a file when told to do so. It reads some of the file's content and tries to figure out what type it is based on the characters found. And as Dave Webb Mentioned the MimeTypes Module under python will work, provided an extension is handy.

Alternatively, if you are sitting on a UNIX box you can use sys.popen('file -i ' + fileName, mode='r') to grab the MIME type. Windows should have an equivalent command, but I'm unsure as to what it is.

python 3 ref: https://docs.python.org/3.2/library/mimetypes.html

mimetypes.guess_type(url, strict=True) Guess the type of a file based on its filename or URL, given by url. The return value is a tuple (type, encoding) where type is None if the type can’t be guessed (missing or unknown suffix) or a string of the form 'type/subtype', usable for a MIME content-type header.

encoding is None for no encoding or the name of the program used to encode (e.g. compress or gzip). The encoding is suitable for use as a Content-Encoding header, not as a Content-Transfer-Encoding header. The mappings are table driven. Encoding suffixes are case sensitive; type suffixes are first tried case sensitively, then case insensitively.

The optional strict argument is a flag specifying whether the list of known MIME types is limited to only the official types registered with IANA. When strict is True (the default), only the IANA types are supported; when strict is False, some additional non-standard but commonly used MIME types are also recognized.

import mimetypes
print(mimetypes.guess_type("sample.html"))

I try mimetypes library first. If it's not working, I use python-magic libary instead.

import mimetypes
def guess_type(filename, buffer=None):
mimetype, encoding = mimetypes.guess_type(filename)
if mimetype is None:
    try:
        import magic
        if buffer:
            mimetype = magic.from_buffer(buffer, mime=True)
        else:
            mimetype = magic.from_file(filename, mime=True)
    except ImportError:
        pass
return mimetype

I'm surprised that nobody has mentioned it but Pygments is able to make an educated guess about the mime-type of, particularly, text documents.

Pygments is actually a Python syntax highlighting library but is has a method that will make an educated guess about which of 500 supported document types your document is. i.e. c++ vs C# vs Python vs etc

import inspect

def _test(text: str):
    from pygments.lexers import guess_lexer
    lexer = guess_lexer(text)
    mimetype = lexer.mimetypes[0] if lexer.mimetypes else None
    print(mimetype)

if __name__ == "__main__":
    # Set the text to the actual defintion of _test(...) above
    text = inspect.getsource(_test)
    print('Text:')
    print(text)
    print()
    print('Result:')
    _test(text)

Output:

Text:
def _test(text: str):
    from pygments.lexers import guess_lexer
    lexer = guess_lexer(text)
    mimetype = lexer.mimetypes[0] if lexer.mimetypes else None
    print(mimetype)


Result:
text/x-python

Now, it's not perfect, but if you need to be able to tell which of 500 document formats are being used, this is pretty darn useful.

For byte Array type data you can use magic.from_buffer(_byte_array,mime=True)