Plasmids 101: Sequence and Ligation Independent Cloning (SLIC)
Type
Blog Post
...enzyme cloning uses short sticky ends to insert genes into plasmid backbones, base pairing between these...Schematic of SLIC. To start the SLIC cloning process, a gene of interest is PCR amplified to add 5′ and 3′ homology... LIC received an important update, courtesy of Addgene depositor Stephen Elledge. His new method, named...flexible. For one thing, those 5′ overhangs can be generated in several ways. One option, imprecise T4 digestion...work directly on PCR products, with 5′ overhangs generated either by annealing a mixture of products or from...regions from the target vector. 5′ overhangs can be generated by T4 polymerase (left branch); by annealing two...concentrations of insert to succeed. 5′ overhangs are generated in the linearized vector by T4 digestion; then...