St. John's (40-18), the champion of the Big East Tournament, is the No. 3 seed, while VCU (34-24), champs of the Colonial Athletic Association tourney, is seeded fourth.
The Rebels and the Cavaliers met last season in Oxford in the Super Regional with Virginia advancing.
Ole Miss head coach Mike Bianco, taking his ninth UM team into an NCAA Regional in ten seasons, said all talk about playing the Cavaliers again is premature. There is first a matchup with St. John's, which beat No. 7 national seed Louisville twice last week in the Big East tourney.
"We've got to play St. John's," said Bianco, whose team is on the road for the second time in the last three seasons for the NCAA Tournament. "We need to take care of business on Friday."
Bianco said the Rebels knew they'd be traveling. They just didn't know where.
"We knew that probably even before last weekend (in Hoover)," he said. "After the final series (against Auburn), we knew we'd likely go on the road regardless of how we played in Hoover. Now we can start focusing on playing again rather than where we go or who the matchups are."
Drew Pomeranz will start the Friday game for Ole Miss against St. John's, which begins at 8 p.m. ET. The first game of the day is at 4 p.m. ET between UVA and VCU.
Bianco said there was little doubt that he would start his ace, considered one of the country's best, in game one.
"We need to win and we need to move on," he said. "To me to not start your ace would be a mistake."
There was another factor to consider as well. Aaron Barrett is a strong second pitcher for the Rebels, who will likely face the Cavaliers in game two on Saturday.
"We have a lot of confidence in Aaron that he can pitch and win," Bianco said. "To have an advantage, you have to win the first two."
The Red Storm batted .327 this season, a number that impressed Bianco.
"It's a very good St. John's team," Bianco said. "They're obviously a very offensive team."
St. John's beat the University of New Orleans 31-6 in a game early this season. The Red Storm has five players batting over .350 but only one with double-digit home runs. Freshman outfielder Jeremy Baltz has 20 round-trippers.
Freshman right-hander Kyle Hansen is the ace of the staff. At 6-foot-7, Hansen brings an ERA of 3.09 and an 8-1 record into the contest.
This is the 33rd NCAA Tournament appearance for St. John's, which ranks seventh all-time in the country. The appearance is the sixth for head coach Ed Blankmeyer and the third in the past four seasons.
"We're very excited to be back in the NCAA Tournament this season," said Blankmeyer. "This is a strong regional. Virginia, Mississippi and VCU are very good programs, but we feel like we have as good of a chance as anybody to win."
Zach Miller, the fifth-year UM senior infielder, said the Rebels are excited to be playing more baseball in 2010.
"It's a new season," he said. "Each weekend now is a new season. It's a great opportunity, and I hope we can capitalize on it."
The Charlottesville Regional is matched against the Norman Regional for a Super Regional the following weekend. Oklahoma is the top seed there with a 44-15 record. California (29-23), North Carolina (36-20), and Oral Roberts (35-25) round out the field at OU.