Serena Williams was the defending champion in the women's singles tournament at the 2016 Australian Open, and was attempting for the second time to equal Steffi Graf's career record of 22 major singles titles, but lost in the final to Angelique Kerber, 4–6, 6–3, 4–6. Kerber became the first woman to win a singles major title after having saved a match point in the first round, which she did against Misaki Doi. She is also the first German of either sex to win a major since Graf at the 1999 French Open.

The top three seeds (Williams, Simona Halep and Garbiñe Muguruza) were in contention for the world No. 1 ranking at the beginning of the tournament. However, Halep and Muguruza were eliminated in the first and third rounds, respectively, with Williams retaining the top spot at the end of the tournament.

The twelve seeds that lost in the opening round was the most in the opening round of any major tournament since the 32-seed draw was adopted at the 2001 Wimbledon Championships.

Johanna Konta became the first British woman to reach the semi-final of a major since Jo Durie in 1983. Zhang Shuai became the first qualifier to reach the Australian Open quarter-finals since Angélica Gavaldón in 1990.

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