The January 1861 law that created the Virginia Convention required that if it voted to secede then the state's voters had to approve or disapprove in a popular referendum. It took place on Thursday, May 23, 1861. By then opposition to secession had virtually disappeared in eastern Virginia as the state's white men mobilized for war. In some eastern and southern counties the vote for secession was unanimous. In the city of Richmond the vote was 3,682 for secession and 3 against, and in Shenandoah County, in the Shenandoah Valley, where 13 men had voted for Abraham Lincoln in November 1860, only 5 voted against secession in May 1861. In many of the counties west of the Blue Ridge Mountains where voters had elected strong Unionists to the convention in February, they ratified the Ordinance of Secession by large majorities.
The official totals certified by the governor were 125,950 for secession and 20,373 against. From unofficial information not identified by the governor, he estimated that another 2,934 men, mostly in eastern or mountain counties from which official returns were not available, voted for secession, and that 11,761 men, mostly in western and northwestern counties, voted against secession.