All pupils from Battlefield Primary School, in the city's South Side, took part in the Guinness World Record attempt organised by SignHealth, a charity for deaf people.
Around 350 children and 18 members of staff gathered in the assembly hall on Wednesday afternoon for the sign2sing event.
They donned SignHealth scarfs and joined schools from across the country in a bid to break the record.
But they won't find out if they've achieved this until it has been verified.
Principal teacher Catriona Brown, who has been learning sign language with her colleague Sarah Gardyne for four years, organised the event.
Ms Brown said: "The kids had a great time, it was fantastic. They all received a certificate for taking part.
"We managed to break the world record last year, so we're all hoping we've done enough this year."
Steve Powell, chief executive at SignHealth, said: "Hundreds of schools from all over the country, including Glasgow and overseas, registered to take part in our Guinness World Records sign2sing project.
"The schools ranged in size from 30 to 800 pupils, which, from our calculations, means more than 135,000 schoolchildren took part in the event.
"The exact number of participants will be confirmed by Guinness World Records at a later date.
"The previous world record, which we set last year, was 114,277 so we are hoping to have smashed this.
"It has been a truly fantastic event and we're extremely grateful to everyone who took part."
Young people sang and signed a song called sign2sing, which was composed especially for the event.
Everyone who took part was asked to donate £1 to raise money for SignHealth's work to improve the mental and phsyical health and well-being of deaf people.