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Yes, absolutely. Although the mother's participation can strengthen results, her DNA is not usually needed in order to obtain conclusive results.
Under Florida law, paternity testing is a requirement if an individual is seeking child support and there is a dispute as to who may be the father of the child.
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If the father is unavailable, it's possible to get DNA from parents, grandparents, siblings, as well as aunts and uncles to complete a test, since they will share much of the DNA as the alleged father. But getting a DNA sample directly from the father will produce the most accurate result.