This article explains who qualifies for citizenship based on their Slovak ancestry.
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There are two basic routes: The Green option: for individuals with a Certificate of a Slovak Living Abroad, and the Orange option: for individuals without the SLA Certificate. Although both routes have different legal requirements for ancestors and applicants, they substantially overlap. Many people with Slovak heritage will be eligible for citizenship through both routes. However, there are cases where only one of the routes is available to applicants (eg too distant ancestors). If your goal is to gain EU citizenship, you might be eligible for Czech or Hungarian citizenship.
The Orange option concerns individuals whose parents, grandparents or great grand-parents were at some point Czechoslovak citizens and were born in today’s territory of Slovakia. The law does not require that they were citizens or lived in Czechoslovakia their entire life. The fact that they gained citizenship at any point in their life will suffice. If your ancestors do not qualify (eg because they were born outside the territory, or were not Czechoslovak citizens), you can still gain citizenship through a certificate of Slovak Living Abroad.
The Green option exists for individuals with a certificate of Slovak Living Abroad Certificate (SLAC) who can convert it into Slovak citizenship after three years of residency. In order to obtain a SLAC, you have to fulfil different requirements for ancestry. Generally, your direct ancestor must have self-identified as being of Slovak nationality. There is no explicit requirement that your ancestors were citizens, or that they ever lived in Slovakia or Czechoslovakia. But you must have proof of their self-identification, such as through census documents.