It is well established that cold storage results in increased resistant starch and a reduced glycemic index in carbohydrate food. However, the effects of cold storage on the glycemic response of the second meal of cereals remain unclear. The aim of this study was to compare the postprandial glycemic responses between the paired glutinous and non-glutinous grains, either fresh-cooked or refrigerated, after both the first and second meals. In this randomized crossover trial, eighteen healthy female participants consumed eight test meals, each containing 50 g of carbohydrate, including fresh-cooked non-glutinous and glutinous rice, non-glutinous and glutinous millet, and their refrigerated counterparts (4 °C for 24 h). Postprandial blood glucose and insulin were measured at 240 min and 120 min after breakfast. After a standard lunch, the participants’ blood glucose concentrations were measured within 180 min. The rapidly digestible starch (RDS), slowly digestible starch (SDS), and resistant starch (RS) contents of the samples were determined by in vitro enzymatic analysis. Cold-stored non-glutinous rice (CR) and cold-stored non-glutinous millet (CM) had a 24.4% and 29.5% lower incremental area under the curve (iAUCglu) of glucose within 240 min compared to the control (fresh-cooked rice non-glutinous, FR), respectively (p < 0.05). There were no significant differences between either the cold or hot glutinous grains and FR with respect to postprandial glycemic and insulinemic parameters. After a standard lunch, the fresh-cooked non-glutinous millet (FM) achieved a 39.1% lower iAUCglu0–180 compared to the FR (p < 0.05). FM had the highest percentage of SDS (64.8%, p < 0.05) among all grain samples. Refrigeration treatment reduced the glycemic excursion only in non-glutinous grains at the first meal, but the FM instead of CM demonstrated a significant second meal effect.