New England Biolabs, N0460S, Acyclonucleotide Set

Related Categories Nucleotide Solutions Applications DNA Sequencing,, PCR,, PCR FAQ Q: Are substrates terminated with 3' acyclonucleosides resistant to 3'-5' exonuclease activity? A: The 3'-5' exonuclease activity of DNA polymerases will excise 3' acyclonucleosides at the end of DNA substrates. Q: Which DNA polymerases incorporate acyclonucleotides? A: Therminator DNA polymerase has been engineered to efficiently incorporate acyclonucleotides. Other exonuclease deficient (exo-) Family B DNA polymerases including Vent exo-and Deep Vent exo- also incorporate acyclonucleotides less efficiently. Acyclonucleotides are not incorporated efficiently by Taq or Klenow exo- DNA polymerase. Q: Can acyclonucleotides be used in Sanger sequencing? A: Acyclonucleotides are terminators that can substitute for ddNTPs in Sanger sequencing reactions when used with Therminator DNA polymerase (Gardner, A. and Jack, W. (2002) Acyclic and dideoxy terminator preferences denote divergent sugar recognition by archaeon and Taq DNA polymerases. Nuc. Acids Res. PMID: 11788725; Gardner, A.F., Joyce , C.M. and Jack, W.E. (2004) Comparative kinetics of nucleotide analog incorporation by Vent DNA polymerase. J. Biol. Chem. 279, 11834-11842. PMID: 14699133). Q: Can acyclonucleotides be used for sequencing-by-synthesis? A: Acyclonucleotides are non-reversible terminators that cannot be used in workflows that require reversal of termination such as sequencing-by-synthesis.