
Dufner had a golden chance to become the first man in the history of the majors to break 63 after putting his approach at the 18th to 12 feet but, perhaps feeling the weight of history, hit a nervy putt which came up a foot short.
(US unless stated)
-9 J Dufner
-7 A Scott (Aus)
-7 M Kuchar
-7 J Furyk
-6 J Rose (Eng)
-6 H Stenson (Swe)
Click here for collated scores
However, his 63 - which beat the course record of 64 first set by Ben Hogan in 1942 - meant the 36-year-old American ended the day on nine-under and two in front of Masters champion Adam Scott along with US compatriots Matt Kuchar and Jim Furyk.
Dufner set the tone for his amazing afternoon round by holing his approach at the par four second for eagle and added further birdies at four and five to turn in 31.
Further birdies at 11, 13 and 16 took him clear of the field and no-one could reel the Ryder Cup star in.
On a par 70 Oak Hill course softened up by morning rain, the greens became dart boards with Justin Rose shooting 29 on the front nine and Peter Hanson firing a 30 on the back. Like Dufner, Webb Simpson also had a 62 in his sights but played his final three holes in one-over for a 64.
Rose's birdie blitz on his second nine - he'd teed off at 10 - hauled him from even par to six-under and very much in the hunt to win his second major in two months following his US Open victory at Merion.
He's the leading European after 36 holes alongside Open runner-up Henrik Stenson, who continued his great form by matching Rose's 66 to finish the day three back in tied fifth. Germany's Martin Kaymer is a further stroke in arrears after a second 68.

Tiger Woods' hopes of ending his five-year wait for a major look very much in the balance though after a patchy second round left him 10 shots behind Dufner.
Needing a low round to get himself into contention after a disappointing first day 71, Woods could never build any momentum and mixed four birdies with four bogeys, including one at the last, for a level par 70.
Defending champion Rory McIlroy will go into the weekend nine shots back although he'll be glad to at least have the chance to retain his trophy.
The Northern Irishman, who won this event by eight shots at Kiawah Island last year, looked to be heading for a missed cut after dropping five shots in five holes to slump to four-over but birdied four of his final six to get back to even par.
Lee Westwood went the other way after, at one point, holding a share of the lead with Scott.
The Englishman, a morning starter, reached six-under after birdies at 12 and 14 but finished at one-under. Bogeys at 17 and 2 halted his momentum before he finished double bogey, bogey to leave his hopes of an elusive first major looking bleak.
Open champion Phil Mickelson has even more work to do after firing a second successive 71 to finish at two-over.
The cut fell at four-over. Notable halfway casualties included Luke Donald along with major winners Ernie Els, Bubba Watson, Charl Schwartzel and Padraig Harrington.