44 cars will attempt to qualify for the 2021 Daytona 500, which will result in four cars being sent home.
| # | Driver | Car | Team |
|---|---|---|---|
| 00 |  Quin Houff | Chevrolet Camaro | StarCom Racing |
| 1 | Kurt Busch | Chevrolet Camaro | Chip Ganassi Racing |
| 2 | Brad Keselowski | Ford Mustang GT | Team Penske |
| 3 | Austin Dillon | Chevrolet Camaro | Richard Childress Racing |
| 4 | Kevin Harvick | Ford Mustang GT | Stewart-Haas Racing |
| 5 | Kyle Larson | Chevrolet Camaro | Hendrick Motorsports |
| 6 | Ryan Newman | Chevrolet Camaro | Roush Fenway Racing |
| 7 | Corey Lajoie | Chevrolet Camaro | Spire Motorsports |
| 8 | Tyler Reddick | Chevrolet Camaro | Richard Childress Racing |
| 9 | Chase Elliott | Chevrolet Camaro | Hendrick Motorsports |
| 10 | Aric Almirola | Ford Mustang GT | Stewart-Haas Racing |
| 11 | Denny Hamlin | Toyota Camry | Joe Gibbs Racing |
| 12 | Ryan Blaney | Ford Mustang GT | Team Penske |
| 13 | Garrett Smithley* | Ford Mustang GT | MBM Motorsports |
| 14 | Chase Briscoe | Ford Mustang GT | Stewart-Haas Racing |
| 15 | Derrike Cope | Chevrolet Camaro | Rick Ware Racing |
| 16 | Kaz Grala* | Chevrolet Camaro | Kaulig Racing |
| 17 | Chris Buescher | Ford Mustang GT | Roush Fenway Racing |
| 18 | Kyle Busch | Toyota Camry | Joe Gibbs Racing |
| 19 | Martin Truex Jr. | Toyota Camry | Joe Gibbs Racing |
| 20 | Christopher Bell | Toyota Camry | Joe Gibbs Racing |
| 21 | Matt DiBenedetto | Ford Mustang GT | Wood Brothers Racing |
| 22 | Joey Logano | Ford Mustang GT | Team Penske |
| 23 | Bubba Wallace | Toyota Camry | 23XI Racing |
| 24 | William Byron | Chevrolet Camaro | Hendrick Motorsports |
| 33 | Austin Cindric* | Ford Mustang GT | Team Penske |
| 34 | Michael McDowell | Ford Mustang GT | Front Row Motorsports |
| 36 | David Ragan* | Ford Mustang GT | Front Row Motorsports |
| 37 | Ryan Preece* | Chevrolet Camaro | JTG Daugherty Racing |
| 38 | Anthony Alfredo | Ford Mustang GT | Front Row Motorsports |
| 41 | Cole Custer | Ford Mustang GT | Stewart-Haas Racing |
| 42 | Ross Chastain | Chevrolet Camaro | Chip Ganassi Racing |
| 43 | Erik Jones | Chevrolet Camaro | Richard Petty Motorsports |
| 47 | Ricky Stenhouse Jr. | Chevrolet Camaro | JTG Daugherty Racing |
| 48 | Alex Bowman | Chevrolet Camaro | Hendrick Motorsports |
| 51 | Cody Ware | Chevrolet Camaro | Rick Ware Racing |
| 52 | Josh Bilicki | Ford Mustang GT | Rick Ware Racing |
| 53 | Joey Gase | Ford Mustang GT | Rick Ware Racing |
| 62 | Noah Gragson* | Chevrolet Camaro | Beard Motorsports |
| 66 | Timmy Hill* | Ford Mustang GT | MBM Motorsports |
| 77 | Jamie McMurray | Chevrolet Camaro | Spire Motorsports |
| 78 | B.J. McLeod | Ford Mustang GT | Live Fast Motorsports |
| 96 | Ty Dillon* | Toyota Camry | Marty Gaunt |
| 99 |  Daniel Suarez | Chevrolet Camaro | TrackHouse Racing |
*Is not locked into the race
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