Training and clinical preparations are underway at it's first site in Halifax, which is due to open later this week.
It will be administering the Oxford-AstraZeneca vaccine and operate from purpose-built facilities in its pharmacy in the town.
Two additional vaccination sites in Huddersfield and Gloucester will also open in the near future, and discussions are underway about the potential to expand to more mass vaccination sites in the coming months.
Boots says it has "extensive knowledge, experience and proven capability in vaccination services making it a perfect partner to support the NHS" in delivering the vaccine across the UK.
It has set up specialist centres in its pharmacies which allow the Covid-19 vaccine to be administered by pharmacists and ensures that exceptional safety, clinical and operational standards are met while achieving the vaccination of at least 1,000 patients per site each week.
Patients will be provided the option to book their vaccination appointment through the NHS booking system, once it is open.
It comes as Health Secretary Matt Hancock said that every adult in the UK will have been offered a Covid-19 vaccination by autumn this year and mass hubs open across the country.
Seb James, managing director of Boots UK, said: “Boots has already conducted over two million COVID-19 tests on behalf of the NHS and now we are proud to play our part in vaccinating the nation.
"Our vaccination sites are clean, clinical environments, in accessible, convenient locations, like high streets, and we have extensive experience in vaccination services, having carried out over a million flu vaccinations in this winter flu season alone.
"We hope to help the NHS and enable more people to get the vaccination quickly. I am incredibly thankful for the whole team’s support in helping make this happen.”
A spokesperson added: “We have worked closely with the Clinical Commissioning Groups (CCGs) to initially open three COVID-19 vaccination sites, based in Halifax, Huddersfield and Gloucester.
"These initial stores will provide rapid experience and feedback to inform a wider roll-out at speed and scale.”