The candidates were asked to put aside prepared campaign slogans Michele Bachmann and Tim Pawlenty traded barbs, and others touted their experience as governors.
Meanwhile on Thursday, Texas Governor Rick Perry said he would join the 2012 nomination race.
Mr Perry was expected to announce his candidacy in South Carolina on Saturday, reports said. A spokesman said Mr Perry would make his intentions "very clear".
The news came hours before the eight Republican rivals began the third debate of the 2012 race, days before most of his rivals take part in the Iowa straw poll in Ames.
The candidates hope to win the Republican nomination to face President Barack Obama in the November 2012 general election.
The Republican field has been slow to take shape this year, although the first balloting will not be held until February in Iowa.
Challenges In Iowa, Ms Bachmann, Tim Pawlenty, Jon Huntsman and others are facing a key test of their ability to emerge as Mr Romney's chief challenger, analysts say.
Mitt Romney, right, has thus far declined to engage his rivals for the nomination on the campaign trail Mr Romney, a former financier and Massachusetts governor, touted his experience in the private sector but was forced to defend his record as governor over similarities between his healthcare policy in Massachusetts and Mr Obama's 2010 law.
Mr Huntsman touted his record as governor of Utah, Mr Pawlenty his record of financial management as Minnesota governor and Ms Bachmann - a Minnesota congresswoman - her opposition to the Democratic legislative agenda.
Mr Romney, who failed in his 2008 bid for the Republican nomination, leads the current pack in polling and fundraising.
Much of the early debate centred on exchanges of attacks between Mr Pawlenty and Ms Bachmann, who are vying to pull to the front of the pack and take on Mr Romney.
Mr Pawlenty repeatedly criticised Ms Bachmann's lack of substantive success in enacting a legislative agenda, and accused her of "making false statements".
"She speaks of leading these efforts in Washington and Minnesota," he said. "Leading and failing is not the objective."
Ms Bachmann, meanwhile, said she opposed a law passed last week that raised America's debt limit.
In addition, pizza restaurant magnate Herman Cain said Islamic "sharia law" should never be recognised in US courts and on immigration, said "America can be a nation of high fences and wide open doors."
Former Pennsylvania Senator Rick Santorum called for a 0% tax on manufacturing, former House Speaker Newt Gingrich attacked "Mickey Mouse questions" from the news media on reports of staff defections in his campaign, and Ms Bachmann said Americans should be allowed to purchase whatever lightbulbs they wanted.
Ms Bachmann was seen as a favourite among the social conservative voters who are influential in the Iowa Republican electorate.
She has earned a name for herself for her fiery conservative rhetoric, though at the last debate in June she impressed with her calm and steady demeanour.
Analysts say her challenge will be to meet the high bar she has set for herself and to continue to show she has a substantive command of policy.
And she may face questions about recent reports a counselling centre her husband runs received government aid and engaged in therapy practices aimed at turning homosexuals into heterosexuals.
Straw poll Mr Huntsman, former Utah Governor and ambassador to China under Mr Obama, has thus far failed to live up to the excitement his entrance into the race generated.
Known as a political moderate, he has pledged to run a civil campaign. Analysts say he needs a breakout performance on Thursday to re-engage his fundraising and support network.
Likewise, Mr Pawlenty, the former Minnesota Governor, needs to regain standing lost since he failed to challenge Mr Romney on healthcare policy during the last debate.
The Ames debate comes two days before an early non-binding "straw poll" contest at the Iowa state fair.
The poll is seen as an early test of support, fundraising and organising prowess.
Among major candidates who have formally entered the race, Mr Romney and Mr Huntsman are not competing in the straw poll.
Also, former Alaska Governor and 2008 Republican vice-presidential nominee Sarah Palin, who has flirted with a candidacy but has not entered the race, has announced a bus tour of the state beginning on Friday.
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