Quick & Easy Site-Directed Mutagenesis Kit

Protocol Overview

This kit's protocol is extremely user-friendly. To begin, you design primers to synthesize your point mutation of interest. The kit includes free access to a surprisingly user-friendly online primer design tool that will design primers for you after inputting your DNA sequence and indicating the specific mutation required. Once the primers have been designed and synthesized, you run a PCR reaction to synthesize the mutation-containing DNA, and then digest the parental DNA with DpnI. Together, those two steps take around 90 minutes. Following DpnI digestion, part of the resulting mutation-containing DNA is transformed into competent cells and plated to achieve individual clones for screening. Determining the presence of your point mutation in each colony involves isolating the DNA and sequencing across your point mutation with PCR primers that flank the region of interest.