Thermo Fisher, K1231, CloneJET PCR Cloning Kit, 20 Reactions

CloneJET PCR Cloning Kit, 20 Reactions is designed for rapid cloning of PCR-amplified DNA and other DNA fragments into a positive-selection plasmid vector. The kit contains the pJET1.2/blunt cloning vector and reagents for 20 ligation reactions. Blunt-ended DNA generated by a proofreading polymerase can be ligated directly; sticky-ended or non-proofreading-polymerase PCR products can be processed with the included DNA Blunting Enzyme before ligation. The vector contains a lethal restriction-enzyme gene interrupted when a DNA insert is ligated into the cloning site, favoring the survival of recombinant clones after transformation. The ligation step can be completed in approximately five minutes, and the system is suitable for both phosphorylated and non-phosphorylated DNA fragments. The multiple-cloning-site arrangement places BglII sites around the insert and also includes a T7 promoter for downstream analysis and transcription applications. The kit can be used with common laboratory E. coli strains for transformation, but competent cells are not part of this assay format. This configuration provides a direct route from a PCR product to a plasmid construct with reduced background from empty-vector transformants.