Scientists have failed to prove remains uncovered in the US belong to explorer Bartholomew Gosnold born in Suffolk.
Scientists still hope to establish remains were Gosnold's
Gosnold helped set up the first permanent English-speaking colony in the New World in 1607 and is ranked among America's founding fathers.
Archaeologists based proof on DNA taken from a body believed to be his sister, Elizabeth Tilney, buried at All Saints Church, Shelley, near Ipswich.
It has now emerged tests revealed that the woman was not Gosnold's sister.
Experts were given special permission by Church of England officials to carry out checks on the remains which had been buried in Shelley for 400 years.
Scientific dead end
A church spokesman said it was unlikely that any further work would be done in the churchyard.
Experts are now to examine other avenues to prove their theory.
"British and American archaeologists were surprised and disappointed to learn that DNA material taken from the remains of a woman buried in Shelley church, Suffolk, turned out to be a scientific dead end," said the church spokesman.
"Nevertheless, they remain confident that remains of a 17th Century captain discovered in Jamestown, Virginia, are those of the leader of the expedition who established the first permanent English settlement in the New World in 1607."