A-Tailing (single nucleotide) with Terminal Transferase

1. Set-up the reaction as follows:

  • PCR-amplified DNA – X
  • 10X Terminal Transferase Reaction Buffer – 7.5ul
  • 1 mM ddATP – 1.5 ul
  • CoCl2 – 7.5 ul
  • Terminal Transferase – 6 ul
  • H2O – X
Total Reaction Volume – 75 ul (This volume can be adjusted based on the volume of PCR-amplified DNA that needs to be added).

2. Incubate the reaction at 37 oC for 1.5 hr.