Home FAQs Can I use my own specific primers for my RCA step in the EnClose™ Cell-free dbDNA™ Synthesis protocol (NEB #E9301) instead of the provided primers?

FAQ: Can I use my own specific primers for my RCA step in the EnClose™ Cell-free dbDNA™ Synthesis protocol (NEB #E9301) instead of the provided primers?

Yes, different specific primers may be used in the RCA reaction specific for the amplification of your sequence of interest. Based on limited observations, primers of 13-16 bases are more efficient than longer primers. As a general primer design rule (RCA, PCR, etc.), try to avoid more than 3 Gs or Cs in the last five bases to avoid mis-priming. It is critical that the 3′ end is protected from the strong exonuclease activity of phi29-XT DNA Polymerase by including a minimum of two phosphorothioate bonds.