HomeFAQsWhy do I not see my PCR product after using the Q5® Site-Directed Mutagenesis Kit?
FAQ: Why do I not see my PCR product after using the Q5® Site-Directed Mutagenesis Kit?
Correct primer design and annealing temperature determination are very important. Use of the NEB online site-directed mutagenesis primer design tool, NEBaseChanger, for design and annealing temperature determination is highly recommended. In addition, ensure that 1) the plasmid template concentration is between 1 and 25 ng, 2) the final primer concentration is 0.5 µM, and 3) the extension time is 20-30 seconds per kb of plasmid.