Shown on commercial television, the ad features three mothers voicing their concerns about how changing the Marriage Act will affect what their children are taught in schools.
"The school told my son he could wear a dress next year if he felt like it .. "
"When same sex marriage laws pass overseas this sort of program becomes widespread and compulsory .. "
"Kids in year seven are being asked to role play being in a same sex relationship .. you can say no."
But less than 24 hours later, several of those claims have been refuted.
The high school at the centre of the claim that a male student was told he could 'wear a dress next year' has told SBS that simply isn't true.
Principal of Frankston High School in Melbourne's east, John Albiston, has called the statement a complete fabrication.
"We've spoken with all the teachers of her son's classes and we know that didn't happen and never did she raise that as a concern when I've met and spoken with her, which I do find interesting."
The ad was commissioned by the Coalition for Marriage, a group led by the Australian Christian Lobby.
The Coalition's David Van Gend has defended the claims, refering to material distrubuted by an LGBTI youth organisation that supports gender-neutral school uniforms.
"This is the sort of gender-bending material being inflicted on our kids and it's confusing them. And that's going to be exponentially worse if we bend gender within the Marriage Act."
He says legalising same-sex marriage will lead to sweeping reforms about how the issue will be taught in schools.
However those claims have been rejected by the Federal Education Minister, Simon Birmingham.
"It is patently ridiculous to suggest that allowing same-sex couples to marry is somehow going to see some new wave of teaching reform sweep across the country, that's just not going to happen. This is a simple issue, and it should not be conflated with other issues."
The ad was also slammed at the launch of a group calling itself Australian Christians for Marriage Equality.
Anglican reverend Keith Mascord has accused the Coalition for Marriage group of scaremongering.
"To put out an ad that links same sex marriage or marriage equality with the safe schools program is, for me, an admission of failure. It's not looking at the substance of the debate, it's fear-mongering. It's irresponsible and wrong."
His comments are supported by the rector of Melbourne's Xavier College, Father Chris Middleton.
Father Middleton is urging parents to consider all sides of the argument before voting, and called upon the church to reflect on the overwhelming support for marriage equality among young people.
"I think my first intention was, 'let's try and lift the debate and see where both sides are coming from'. Personally I don't think the case has been made against same sex marriage. So I'd be leaning towards 'yes'."
Opposition leader Bill Shorten has called the ad offensive and hurtful to LGBTI Australians and their families.
"When I first saw the ad I wondered if it was send-up. I thought it was so primitive and thought it was rubbish. The idea that if you vote in this $122 million postal survey that somehow your little boys will go to school and be made to wear dresses it's just ridiculous. It's offensive."
However Federal Health Minister Greg Hunt says Labor rejected the chance for strict advertising standards to be in place when it rejected a plebiscite, and only has it itself to blame for this sort of advertising in the lead up to the postal vote.
"We offered the ALP a chance to have every single protection that they wanted and they rejected it. They rejected a legislated plebiscite. So I'll leave that to Mr Shorten to explain why he rejected the protections."
