Methods for amplification and sequencing using thermostable TthPrimPol

A method for replicating, amplifying or sequencing of DNA in the absence of a primer comprising the following steps

• a) providing
• i) a protein that has both polymerase and primase activity, and has a sequence that is at least 80% identical to SEQ ID NO:
• 1, or
• ii) a conjugate comprising a protein moiety that has both polymerase and primase activity, wherein the protein moiety of the conjugate has a sequence that is at least 80% identical to SEQ ID NO:
• 1,
• b) providing a template nucleic acid,
• c) providing nucleotides and/or nucleotide derivatives for incorporation in a complementary strand of nucleic acids,
• d) providing a suitable buffer,
• e) providing a second polymerase, and
• f) contacting the materials of steps a-e in the absence of a primer and without the presence of a protein having primase activity other than the protein or conjugate provided in step (a) for a suitable amount of time to replicate, amplify or sequence the template nucleic acid.