Under the hood

This section documents some details about what happens when a BedTool object is created and exactly what happens when a BEDTools command is called. It’s mostly useful for developers or for debugging.

There are three kinds of sources/sinks for BedTool objects:

  • filename
  • open file object
  • iterator of Interval objects

Iterator “protocol”

BedTool objects yield an Interval object on each next() call. Where this Interval comes from depends on how the BedTool was created and what format the underlying data are in, as follows.


If BED/GTF/GFF/VCF format, then use an IntervalFile object for Cython/C++ speed.

If SAM format, then use an IntervalIterator. This is a Cython object that reads individual lines and passes them to create_interval_from_list, a Cython function. create_interval_from_list does a lot of the work to figure out what format the line is, and this is how we are able to support SAM Interval objects.

If BAM format, then first do a Popen call to samtools view, and create an IntervalIterator from subprocess.PIPE similar to SAM format.

Open file-based

All formats are passed to an IntervalIterator, which reads one line at a time and yields an Interval object.

If it’s a BAM file (specifically, a detected bgzip stream), then it’s actually first sent to the stdin of a samtools Popen call, and then the subprocess.PIPE from that Popen’s stdout is sent to an IntervalIterator.

Iterator or generator-based

If it’s neither of the above, then the assumption is that it’s already an iterable of Interval objects. This is the case if a BedTool is created with something like:

a = pybedtools.example_bedtool('a.bed')
b = pybedtools.BedTool((i for i in a))

In this case, the (i for i in a) creates a generator of intervals from an IntervalFile – since a is a filename-based BedTool. Since the first argument to the BedTool constructor is neither a filename nor an open file, the new BedTool b’s .fn attribute is directly set to this generator … so we have a generator-based BedTool.

Calling BEDTools programs

Depending on the type of BedTool (filename, open file, or iterator), the method of calling BEDTools programs differs.

In all cases, BEDTools commands are called via a subprocess.Popen call (hereafter called “the Popen” for convenience). Depending on the type of BedTool objects being operated on, the Popen will be passed different objects as stdin and/or stdout.

In general, using a filename as input is the most straightforward – nothing is passed to the Popen’s stdin because the filenames are embedded in the BEDTools command.

Using non-filename-based BedTools means that they are passed, one line at a time, to the stdin of the Popen. The commands for the BEDTools call will specify “stdin” in these cases, as is standard for the BEDTools suite.

The default is for the output to be file-based. In this case, an open tempfile object is provided as the Popen’s stdout.

If the returned BedTool is requested to be a “streaming” BedTool, then the Popen’s stdout will be subprocess.PIPE, and the new BedTool object will be open-file based (which is what subprocess.PIPE acts like).

Specifically, here is the information flow of stdin/stdout for various interconversions of BedTool types … .

filename -> filename:

The calling BedTool is filename-based and stream=False.

  • stdin: None (the filenames are provided in the BEDTools command)
  • stdout: open tempfile object
  • new BedTool: filename-based BedTool pointing to the tempfile’s filename
filename -> open file object:

The calling BedTool is filename-based and stream=True is requested.

  • stdin: None (provided in the cmds)
  • stdout: open file object – specifically, subprocess.PIPE
  • new BedTool: iterator-based BedTool. Each next() call retrieves the next line in subprocess.PIPE
open file object -> filename:

The calling BedTool is from, e.g., subprocess.PIPE and there’s a saveas() call to “render” to file.

  • stdin: each line in the open file object is written to subprocess.PIPE
  • stdout: open file object – either a tempfile or new file created from supplied filename
  • new BedTool: filename-based BedTool
open file object -> iterator:

The calling BedTool is usually based on subprocess.PIPE, and the output will also come from subprocess.PIPE.

  • stdin: each line from the open file is written to subprocess.PIPE
  • stdout: open file object, subprocess.PIPE
  • new BedTool: filename based on subprocess.PIPE