| Rank | City | 2000 Population |
|---|---|---|
| 1 | Collinwood | 1,022 |
| 2 | Medina | 987 |
| 3 | Piperton | 982 |
| 4 | Cornersville | 977 |
| 5 | Luttrell | 917 |
| 6 | Huntland | 913 |
| 7 | Gates | 899 |
| 8 | Alexandria | 881 |
| 9 | Decaturville | 856 |
| 10 | Rockford | 800 |
| 11 | Gallaway | 740 |
| 12 | Trimble | 728 |
| 13 | Palmer | 714 |
| 14 | Maury City | 705 |
| 15 | Berry Hill | 676 |
| 16 | Puryear | 667 |
| 17 | Orlinda | 612 |
| 18 | Stanton | 615 |
| 19 | Petersburg | 581 |
| 20 | Sunbright | 578 |
| 21 | Adams | 569 |
| 22 | Beersheba Springs | 553 |
| 23 | Wartrace | 551 |
| 24 | Gadsden | 550 |
| 25 | Copperhill | 511 |
| 26 | Baileyton | 506 |
| 27 | Burlison | 453 |
| 28 | Sardis | 445 |
| 29 | Rossville | 400 |
| 30 | Iron City | 368 |
| 31 | Liberty | 368 |
| 32 | Ramer | 354 |
| 33 | Finger | 351 |
| 34 | Saltillo | 342 |
| 35 | Hornsby | 306 |
| 36 | Yorkville | 293 |
| 37 | Milledgeville | 287 |
| 38 | Centertown | 258 |
| 39 | Parkers Crossroads | 242 |
| 40 | Oakdale | 240 |
| 41 | Slayden | 207 |
| 42 | Medon | 191 |
| 43 | Hickory Valley | 136 |
| 44 | Orme | 124 |
| 45 | Shelbyville | 16,342 |
| 46 | Greeneville | 15,390 |
| 47 | Mount Juliet | 14,172 |
| 48 | Clarksville | 103,944 |
| 49 | Brownsville | 10,766 |
| 50 | Humboldt | 9,464 |
| 51 | Clinton | 9,460 |
| 52 | Portland | 8,642 |
| 53 | Spring Hill | 8,251 |
| 54 | White House | 7,493 |
| 55 | Lakeland | 7,228 |
| 56 | Winchester | 7,359 |
| 57 | Harriman | 6,771 |
| 58 | Dayton | 6,522 |
| 59 | Murfreesboro | 70,858 |
| 60 | Church Hill | 5,954 |
| 61 | Lynchburg, Moore County | 5,765 |
| 62 | Millersville | 5,472 |
| 63 | McKenzie | 5,310 |
| 64 | Kingston | 5,309 |
| 65 | Trenton | 4,704 |
| 66 | Selmer | 4,524 |
| 67 | Oak Hill | 4,502 |
| 68 | Huntingdon | 4,373 |
| 69 | Jackson | 59,922 |
| 70 | Madisonville | 4,006 |
| 71 | Smithville | 3,995 |
| 72 | Atoka | 3,772 |
| 73 | Centerville | 3,823 |
| 74 | Gatlinburg | 3,444 |
| 75 | South Pittsburg | 3,280 |
| 76 | Dresden | 3,072 |
| 77 | Monterey | 2,761 |
| 78 | Arlington | 2,633 |
| 79 | Parsons | 2,546 |
| 80 | Mountain City | 2,494 |
| 81 | Jellico | 2,452 |
| 82 | Lakewood | 2,348 |
| 83 | Alamo | 2,400 |
| 84 | Halls | 2,317 |
| 85 | Franklin | 46,376 |
| 86 | Caryville | 2,258 |
| 87 | Carthage | 2,254 |
| 88 | Estill Springs | 2,214 |
| 89 | Hollow Rock | 960 |
| 90 | Coalmont | 949 |
| 91 | Kingsport | 45,383 |
| 92 | White Bluff | 2,154 |
| 93 | Byrdstown | 900 |
| 94 | Dandridge | 2,116 |
| 95 | Crab Orchard | 842 |
| 96 | St. Joseph | 830 |
| 97 | New Johnsonville | 1,904 |
| 98 | Lake City | 1,883 |
| 99 | Jamestown | 1,838 |
| 100 | New Hope | 1,040 |
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