Ek/lic cloning manual

2. The vector is compatible with Novagen Ek/LIC Cloning Kits, which allows you to move the gene into the LIC vectors from Novagen if needed. 3. The vector is compatible with Invitrogen Gateway® system, which allows you to move the gene into other expression vectors from Invitrogen if needed. 4. Ligation Independent Cloning (LIC) Ligation Independent Cloning (LIC) is a technique developed in the early s as an alternative to restriction enzyme/ligase cloning. Inserts are usually PCR amplified and vectors are made linear either by restriction enzyme digestion or by PCR. This creative technique uses the 3’ → 5’ exo activity of. Description; Overview: The pET Xa/LIC vector is an Xa/LIC version of pETb(+). It is designed for cloning and high-level expression of target proteins fused with the aa Trx•Tag™ thioredoxin protein, and His•Tag ® and S ® Tag™ coding sequences that are cleavable with Factor Xa protease. The plasmid contains a strong T7lac promoter, an optimized RBS, the coding sequence for the.

directional LIC cloning technology to streamline and facilitate the process of insert cloning into the expression vector. • 95% clones with desired insert • Directional cloning • 15 minutes procedure • Restriction digestion and ligation is not involved aLICator LIC Cloning Kits Features aLICator LIC Cloning and Expression Kits. protein purification. All Ek/LIC vectors possess the same Ek/LIC cloning site; thus, the same Ek/LIC-prepared target insert can be annealed into any of the Ek/LIC vectors. The LIC method uses the 3'→5' exonuclease activity of T4 DNA Polymerase to create specific or base single-stranded overhangs in the Ek/LIC vector. Cloning strategy for LIC Duet™ Adaptors. After ORF 1 and ORF 2 are amplified with primers that include the indicated 5'-LIC extension, the PCR inserts are treated with LIC-qualified T4 DNA Polymerase (+dATP), annealed to the LIC Duet Adaptor and Ek/LIC vector, and transformed into competent E. coli. Components.

The pET Ek/LIC vector is prepared for rapid, directional cloning of PCR-amplified DNA for high-level expression of polypeptides. Using specifically designed primers for amplification and the pET Ek/LIC Cloning Kit (Cat. No. ), inserts can be efficiently cloned without the need for restriction digestion or ligation. protein purification. All Ek/LIC vectors possess the same Ek/LIC cloning site; thus, the same Ek/LIC-prepared target insert can be annealed into any of the Ek/LIC vectors. The LIC method uses the 3'→5' exonuclease activity of T4 DNA Polymerase to create specific or base single-stranded overhangs in the Ek/LIC vector. The pET Ek/LIC vector is an Ek/LIC version of pETb(+). It is designed for cloning and high-level expression of target proteins fused with the His•Tag ® and S ® Tag™ coding sequences that are cleavable with enterokinase (Ek) protease.