The station is on the Great Western Main Line, 31 miles 1 chain (31.01 mi; 49.9 km) west of London Paddington.

[1] It is the junction station for the Henley-on-Thames branch and is served by local services operated by Great Western Railway and Elizabeth line.

The first Twyford station opened on 1 July 1839 and was the terminus of the GWR until 30 March 1840, pending the completion of Sonning Cutting.

The station was completely reconstructed into the form largely visible today, with new platforms (1 and 2) to serve the fast lines and a new footbridge.

A cattle dock and coal yard were built opposite the Henley bay (platform 5) and a weighbridge provided which can still be seen (outside what is now the office of a taxi company).

The new track layout was much more complex and required the construction of two signal boxes, East and West, to replace the original one which stood on the up platform by the footbridge steps.

The GWR was nationalised on 1 January 1948, becoming part of the Western Region of British Railways, but apart from new signs this had little effect at Twyford until the 1960s.

In 1975 the road bridge was reconstructed and platforms 1 and 2 altered to reduce the curve through the station and make the main lines suitable for 125 miles per hour (201 km/h) High Speed Trains.

The GWR pagoda cycle shed was removed from platform 4 at this time and moved to the garden of the former Station Master's house, where it can still be seen.