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A single cylinder Yamaha research engine was operated with gasoline HCCI combustion using negative valve overlap (NVO). The injection strategy for this study involved using fuel injected directly into the cylinder during the NVO period (pre-DI) along with a secondary injection either in the intake port (PI) or directly into the cylinder (DI). The effects of timing of the pre-DI injection along with the percent of fuel injected during the pre-DI injection were studied in two sets of experiments using secondary PI and DI injections in separate experiments. Results have shown that by varying the pre-DI timing and pre-DI percent the main HCCI combustion timing can be influenced as a result of varied heat release during the negative valve overlap period along with hypothesized varied degrees of reformation of the pre-DI injected fuel. In addition to varying the main combustion timing the ISFC, emissions and combustion stability are all influenced by changes in pre-DI timing and percent.