User:Mbennie/Notebook/Papers and Books/Methods

PCR-mutagenesis-restenzymes pmid=15659713
 * Uses type II restriction enzymes to rapidly induce desired mutations into DNA
 * Designs two primers around mutation site with type II (Ear1, BsmA1, BsmB1, Sap1, Ksp6321, BpuA1, Alw261, Eam11041) restriction enzyme sites
 * Performs PCR to create two strands of DNA with mutation and restriction enzyme sites (100ng of template, 1 uM primer pair, 200 uM dNTP, 2.5 U of Taq polymerase)
 * Cuts DNA with restriction enzymes and ligates back together (10 U enzyme, 400 U T4 DNA ligase)
 * Performs gel purification to get desired product
 * Cuts and ligates into a plasmid