I have just installed Visual Studio 2013, created an MVC Web Application project and noticed a new file in the project template called Startup.cs.
What is this, how is this different from Global.asax.cs and are there any good best practices on what to use this for?
Every OWIN application has a startup class where you specify components for the application pipeline.
If you start a new Visual Studio project, you'll see pieces of OWIN in it. OWIN is a specification that defines an API for framework and servers to cooperation. The point of OWIN is to decouple server and application. For example, ASP.NET Identity uses OWIN security, SignalR self hosting uses OWIN hosting, and etc., the examples all use OWIN, therefore they all need to have a startup class, that is defined in "Startup.cs" file.
The Global.asax, the ASP.NET application file, is an optional file that contains code for responding to application-level events raised by ASP.NET or by HttpModules.
For more details:
OWIN
http://www.asp.net/aspnet/overview/owin-and-katana
Global.asax
http://msdn.microsoft.com/en-us/library/1xaas8a2(v=vs.71).aspx
You can find more ideas about why OWIN in the following article:
http://www.asp.net/aspnet/overview/owin-and-katana/an-overview-of-project-katana
The Startup class is the convention that Katana/OWIN looks for to initialize the pipeline. When your app starts, the code inside of the Configuration function is run to set up the components that'll be used. In the MVC 5 templates, it's used to wire up the authentication middleware which is all built on top of OWIN.
If you want to use dependency injection with OWIN, check out this project on GitHub: DotNetDoodle.Owin.Dependencies
The file seems to be related to SignalR. Quoting the VS 2013 release notes: