APOBEC3A is not recommended for dsDNA substrates. Activity on dsDNA was determined to be less than 10% by incubating 1 µl of APOBEC3A with 5 pmol of C-containing 44-mer dsDNA for 30 min. at 37°C in a total reaction volume of 20 µl in 1X Deamination Reaction Buffer.
For dsDNA substrates, we recommend denaturation of dsDNA to ssDNA before deamination by adding formamide to the dsDNA (in water) and incubating at 85°C for 10 minutes, followed by immediate cooling on ice before proceeding to the APOBEC3A reaction.
Alternatively, dsDNA can be denatured by adding 4 µl of 0.05 N NaOH to dsDNA (in water) and incubating at 85°C for 10 minutes, followed by immediate cooling on ice before proceeding to the APOBEC3A reaction.
