DNA samples recovered from items of clothing are often attributed to the wearer and one or more individuals who may have contacted the item during an alleged criminal activity. Another scenario often proposed by defence counsel is that DNA was transferred from a previously contacted item/surface unrelated to the activity of interest onto the item of clothing. Under such scenarios, DNA may also be transferred from the clothing to the item/surface with which it comes into contact. One such surface is flooring, upon which clothing may be placed while not being worn or may be contacted during wearing, such as falling or being forced to the ground. This study investigates the transfer of DNA to and from clothing and flooring when different contacts are applied between the two surfaces in an environment representative of what investigators would encounter in routine casework, a residential environment. Participants were provided with two sets of new and unused upper and lower garments to wash then wear for ~8 h inside their own home before storing them in paper evidence bags. The two sets of clothing were taken to a home occupied by unrelated individuals, where one set was placed on the floor ('passive') by the researcher while the other was worn by the participant who laid with their back on the floor, rolled to one side and back, then stood up ('active'). Within the houses sampled, the main bedroom was targeted as flooring types and histories of use were more consistent across houses and less variation in DNA profile composition was previously observed for samples collected in the same room. Samples were collected from predetermined areas of the clothing and flooring where contact did and did not occur. Reference profiles were obtained from wearers and individuals they lived with, as well as occupants of the home. DNA transfer was observed from clothing to flooring and from flooring to clothing in both 'active' and 'passive' situations, though greater where a situation involved the application of pressure and friction ('active'), and only where contact between clothing and flooring occurred. Results from this study inform on the composition of DNA profiles one is likely to obtain from an item of clothing or a flooring surface following a similar contact event between the two substrates and will aid investigators when interpreting DNA evidence recovered in a domestic environment and the activities leading to its transfer and subsequent recovery.