I started a new digest. Before I used HSS11 and Eagl Hr enzymes to cut the plasmid. Through the software, I selected Pcil and BspHI enzymes. After cutting the plasmids I blunted the ends with Klenow and dNTPs and began PCR (polymerase chain reaction) purification to isolate the pure DNA strands. I am attempting this digest on my own, although my lab technician still teaches me temperature moderation is important throughout the process. For example, enzymes must be stored at below-freezing temperatures, but incubated at 37C.
