The Cardinals started Holliday at first base on Opening Day of the 2016 season, a position he had not previously played during his career. On May 6 against Pittsburgh, Holliday collected his 512th hit at Busch Stadium, breaking his personal tie of 511 hits at Coors Field. Having already achieved 500 hits at both stadiums, he became the only active player with at least 500 hits at multiple stadiums, and just the fourteenth to do so within the previous 50 years. Holliday hit a home run on May 30 that traveled off Jhan Mariñez and nearly exited Miller Park in a 6–0 defeat of the Brewers. The next day, also against the Brewers, he collected his 1,000th career hit with the Cardinals.