Protesters gathered tyres, tree trunks and rocks to build new roadblocks in Khartoum's northern Bahari district, a witness told AFP, but riot police swiftly moved in and fired tear gas at them.
"Almost all internal roads of Bahari have roadblocks. Protesters are even stopping residents from going to work," said the witness.
The latest bid by demonstrators to close off streets in the capital comes nearly a week after a deadly raid on a sit-in outside army headquarters which left dozens dead.
The bloody crackdown prompted the Sudanese Professionals Association, which first launched protests against longtime ruler Omar al-Bashir in December, to announce a nationwide civil disobedience campaign starting Sunday.
The SPA said the movement will end only after the military rulers, who took over after Bashir's ouster two months ago, transfer power to a civilian government.
Khartoum residents have mostly remained indoors over the past few days and the downtown business district was largely shut on Sunday.
Several vehicles of the feared Rapid Support Forces (RSF), blamed by witnesses for Monday's killings, were seen moving across some parts of the capital loaded with machine guns.
Buses were not running in several districts, but private vehicles were ferrying passengers in some areas.
Airlines have scrapped most of their Sudan flights since the deadly raid and several passengers were left queueing outside Khartoum airport's departures terminal Sunday, although it was unclear whether any flights would take off.
In Khartoum's twin city of Omdurman, across the River Nile, many shops and markets remained closed but residents were seen buying staples in some grocery stores.
"Troops were also seen removing roadblocks from some streets in Omdurman," an onlooker said.
Residents have remained on edge since the raid on the sit-in, which killed at least 115 people according to doctors close to the demonstrators.
The health ministry says 61 people died nationwide in the crackdown, 49 of them by "live ammunition" in Khartoum.
Witnesses say the assault was led by the RSF, who have their origins in the notorious Janjaweed militia, accused of abuses in the Darfur conflict between 2003 and 2004.
Demonstrators had been camped out for weeks in Khartoum to pressure the ruling generals into transferring power, but talks between protest leaders and the military broke down mid-May.
Ethiopian Prime Minister Abiy Ahmed travelled to Sudan on Friday in a bid to revive negotiations, holding separate meetings with the two sides after which he called for a "quick" democratic transition.