In this study, the focus was on the role of non-sulphide gangue (NSG) minerals. It was hypothesised that specific NSG minerals have a detrimental effect on flotation recovery of gold bearing minerals and their separation selectivity. In order to test this hypothesis and delineate the effect of the various gangue minerals, a new approach was taken. This involved first isolating the various mineral components of a double-refractory gold ore from one of the Carlin-type deposits using a gravity separation technique. Then flotation experiments were performed using a mixture design on various mixtures of these isolated components under controlled conditions. The results of these mixture experiments supported the hypothesis and demonstrated, for the first time for these types of ores, that even small amounts of NSG minerals, especially carbonaceous matter and clays, had a large adverse effect on the flotation of sulphides and selectivity of separation. While it is tempting to attribute the observed effects solely to slime coating, there is no basis to do so at this stage; it is more reasonable to propose that multiple contributions exist. The results of this study provide the much-needed context and direction for further fundamental studies and for developing processing strategies.