This is the easiest method to construction of mammalian or E coli expression vectors.
Cloning is often limited by cost, labor-intensity, and incompatibility of restriction enzyme sites. Traditional in vitro cloning requires a variety of end-joining enzymes (e.g. T4 ligase, topoisomerase, blunt-end joining enzymes) or specific strand-exchange recombinases (e.g. CRE, loxP, Gateway BP, LR recombinase). Generating overhanging or blunt ends requires preparing vectors by either restriction enzyme digestion or end polishing, or purchasing specific vectors and recombinases.
The engineered E Coli strain in GenTarget’s EcoTM PCR Cloning Kit has an enhanced homologous recombination function enabling an in vivo joining reaction for cloning with no tube reactions.
GenTarget provides E Coli cloning cells with a selection of built-in vectors for mammalian or E Coli expression systems. A proprietary process for making ready-to-use, E Coli cells with built-in vectors ensures low background and a positive cloning rate of greater than 90%.
The revolutionary Eco Fusion in vivo PCR is the easiest PCR cloning method available:
- Simply amplify your gene of interest with a primer pair that is flanked with short arms homologous to the expression vector
- Add 1µl of purified PCR into the engineered, ready-to-use cloning cells
- Immediately proceed to transformation.
You do not need to buy expression vectors. The expression-optimized E Coli or mammalian expression vector is already built into the cloning cells. You just select the cloning cells with the built-in vector that meets your needs.
For E Coli expression, the verified positive clones can be used directly for T7-based or non T7-based) protein expression, depending on the built-in vector, thereby eliminating the second transformation into an expression strain, such as BL21. [Note: If your target is very toxic, directly cloning it into an expression strain may cause difficulties. For such cases, use our self-controlled PCR cloning cells (Cat: IC-1003) ]
The easy Eco PCR cloning method eliminates the labor intensive and sometimes troublesome vector preparation procedures of other cloning methods. There is no need for any in vitro tube reactions such as ligation, Topo jointing, or In-fusion reaction.
Let E Coli do the job for you In Vivo…and do it directly in the expression strain!
GenTarget provides a series of Eco cloning kits with different built-in vectors. Please check our product line (Eco PCR Cloning Kits) for details.