In this paper, the effect of pre-cooling treatment on the low resistivity C54 phase titanium silicide film growth was investigated. Our experimental results and micro-structural analysis show that, by introducing such cooling treatment into the titanium silicide process to precede the conventional rapid thermal annealing, the low resistivity C54 phase formation can be enhanced. Defects at the Si/Ti interface caused by the thermal mismatch between titanium and silicon layers during the cooling treatment were found to contribute to the increase of the C49 nucleus sites. This help to supply more C49 grain boundaries and triple junction sites at which the C54 phase could nucleate. This discovery has the potential to reduce the complexity and cost associated with forming low resistivity titanium silicide on sub-micron structures for future ULSI application.