Scottish Tory leader Ruth Davidson insisted there will not be a second referendum on independence "anytime soon" as she accused the SNP of "quietly ditching" the policy as a campaign tool in the General Election.
Ms Davidson said she did not accept Nicola Sturgeon's party had a mandate for a fresh ballot to be held as the First Minister wants.
Ms Sturgeon claims there is a "cast iron" mandate for another referendum after her manifesto in 2016 made clear another vote could take place if there was a "material change in circumstances" from 2014, such as Scotland being removed from the European Union against its wishes.
After 62% of Scots voted to keep the UK in Europe, Ms Sturgeon instructed Scottish Government staff to start drawing up the necessary legislation for another vote on the issue.
The First Minister wants to see a second independence referendum take place between autumn 2018 and spring 2019, to give Scots an alternative to Brexit.
But Ms Davidson said the SNP had lost the overall majority they had had at Holyrood in the 2016 election, and said: "I don't accept they do (have a mandate)."
The Scottish Conservative leader told the BBC's Sunday Politics Scotland: "With the current trajectory of support in this country going down for another independence referendum I don't see it happening any time soon, because the people of Scotland don't want it, I'm going to stand up for them.
"There is no support for another independence referendum, it doesn't look like there is going to be any support for another referendum, and as we see from Nicola Sturgeon trying quietly this week to ditch independence as a campaigning tool, we're not going to see it any time soon."
The Conservative manifesto for the General Election makes clear that there should not be another referendum without "public consent" and until the Brexit process has "played out".
Ms Davidson said that meant another vote could not take place "until we know how it's working on the ground".
"Of course there's going to be a primary transfer of powers as we leave the European Union from Brussels to Westminster, but there is also then going to be a secondary transfer of powers to the devolved administrations and actually even to local authorities in England too.
She claimed: "The SNP are going to argue for another referendum until the cows come home, you ask them any question at all and their answer is always independence."
But she stated: "There's two principles here. One, Scotland cannot be dragged back to another independence referendum when we don't know what the options look like because we don't know what Brexit looks like, we don't know what independence looks like because Nicola Sturgeon can't even tell us if we'd be in or out of the EU or EFTA (the European Free Trade Association) or something else.
"Nor should we be dragged back there when the people of Scotland don't want it."
Scottish Labour leader Kezia Dugdale also told the same programme she was "absolutely clear" in her opposition to independence and a second referendum
"I am opposed to independence and a second independence referendum, as indeed is the Labour Party in both the UK and Scottish manifestos. That is clear cut, because of the damage of £15 billion of additional cuts."
Instead she said Labour had a "clear commitment to the presumption of devolution and a radical reformed United Kingdom, so we can reject the extremes of the SNP and hard right wing Tory Brexit".
Ms Dugdale said: "People don't want another referendum. It's the SNP who told us the last referendum was a once in a lifetime, once in a generation opportunity.
"It's very clear that after the EU referendum Nicola Sturgeon used Brexit as an excuse for another referendum, she pointed to her manifesto as the reason for that.
"She said Brexit was an excuse for a second independence referendum but the problem the SNP have now is they can't tell us whether Scotland's gets to stay, whether we have to reapply to the European Union, or whether we're going to have an EFTA or Norway type arrangement.
"Her whole justification, her mandate, her manifesto commitment for a referendum has fallen apart because she can't be clear on Europe."