Cotton (Gossypium hirsutum) is sensitive to cold conditions during germination and establishment. Identifying cultivars with cold tolerance offers opportunities for future breeding efforts and provides growers with increased flexibility for sowing date options. Developing cultivars with chilling tolerance will allow cultivars to withstand the cold shocks during early cotton growth (15th September - 30th November), allowing better establishment under cool conditions, and reducing the costs associated with replanting. This project aims to utilise germination chill protocols, and investigate their applicability in determining genetic variation in seedling chilling tolerance. Seeds of ten cultivars (DP16, Namcala, Pima A-8, Sicala 350B, Sicot 289RR, Sicot 71, Sicot 75, Sicot 81, Siokra V-18 and TL) were germinated at four temperatures (14, 18, 22 and 30) and germination probability (percentage) was determined after 4, 7 and 10 d. Ten seedlings were selected randomly and the length from the hook of the hypocotyl to the tip of the radicle (seedling length) was measured. Laboratory tests were correlated to an early planted (August) field experiment conducted 40 km west of Narromine, central west NSW. Seedling length provided a better indication of cold tolerance than germination probability. The best correlation with field emergence was with the Cool-Warm Seedling Length test (average of seedling length between cool temperature 14°C at Day 7 and warm temperature 30°C at Day 4) which provided an R2 of 0.73. The Cool-Warm Vigour Index (average of seedling germination between cool temperature14°C at Day 7 and warm temperature 30°C at Day 4) also provided a correlation with field emergence only when Namcala (outlier) was removed (R2 = 0.62). The electrolyte leakage test provided a negative correlation with field emergence. The laboratory and field experiments indicated that there was some genetic variation in the selected cotton cultivars, with Namcala, DP16, Sicot 75, Sicala 350B and Siokra V-18 showing some degree of cold tolerance during field emergence and laboratory tests. Further research is needed to test more cultivars to validate the Cool-Warm Seedling Length test (Tuck Test).