Novotná turned professional in 1986. In the early years of her career, she was known primarily for her success as a doubles player. In the early-1990s, Novotná began to have success in singles once four-time Grand Slam singles champion Hana Mandlíková became her coach.
Novotná reached her first Grand Slam singles final in 1991 at the Australian Open, where she lost to Monica Seles 5-7, 6-3, 6-1.
Two years later, Novotná reached her first singles final at Wimbledon, where she faced Steffi Graf. After losing a tight first set, Novotná took a 6-7, 6-1, 4-1, 40-15 lead. With victory seemingly in her grasp, she lost her nerve and began missing easy shots, sometimes hitting the ball out by wide margins (including an infamous overhead smash that hit the back tarp). Graf took the next five games and the title. During the prize presentation ceremony, a distraught Novotná burst into tears and cried on the Duchess of Kent's shoulder. The Duchess comforted her by saying that she was sure Novotná would win the title one day. But at the time, many doubted that this would happen given how dramatically she had choked against Graf.
Olympic medal record Women’s Tennis
- Silver 1988 Seoul Doubles
- Silver 1996 Atlanta Doubles
- Bronze 1996 Atlanta Singles
It took four years for Novotná to reach another Wimbledon final. In 1997, she faced Martina Hingis. Novotná won the first set. But she then succumbed to the Swiss teen's accurate passing shots and lost 2-6, 6-3, 6-3. However, Novotná won the 1997 WTA Tour Championships and finished the year ranked a career-high World No. 2 in singles.
Novotná's moment of Wimbledon glory finally arrived in 1998. After routing a young Venus Williams in a quarterfinal, Novotna avenged the previous year's loss by ousting Hingis in a semifinal and veteran Nathalie Tauziat in the final 6-4, 7-6.
She won 12 Grand Slam women's doubles titles (four at Wimbledon, three at the French Open, three at the U.S. Open, and two at the Australian Open) and 4 Grand Slam mixed doubles titles (two at the Australian Open, one at Wimbledon, and one at the U.S. Open). She was 11 times the year end top ranked doubles player.
Novotná was a member of the Czechoslovakian team that won the Fed Cup in 1988. At the Olympic Games, Novotná was a women's doubles silver medalist in 1988 and 1996 and a singles bronze medalist in 1996.
Novotná retired from the professional tour in 1999. During her 14-year career, she won 100 titles (24 in singles and 76 in doubles).
Novotná was inducted into the International Tennis Hall of Fame in 2005.