FAQ: Why does the HF version of the enzyme have a different recommended buffer than the wild type enzyme?

In many cases, changing the charged amino acids of a restriction endonuclease gene results in changes in buffer preference. These changes can be significant. For example, wild type SalI has a strict requirement for NEBuffer 3, a high ionic strength buffer. However, SalI-HF works well in NEBuffer 2 and NEBuffer 4, which are moderate ionic strength buffers.