Fusion In Vivo PCR Cloning
EcoTM cloning technology
Our proprietary high-throughput Fusion in vivo cloning technology eliminates the labor-intensive and sometimes troublesome vector preparation procedures of other cloning techniques. It does not need any kind of In Vitro tube reaction, such as ligation, Topo joining, or In-fusion reaction.
Let E Coli do the job for you in vivo…and do it directly in the expression strain!
With this technology the same PCR product can be used to make both Gateway-compatible Entry and DEST/expression clones without the use of BP or LR enzymes.
GenTarget’s revolutionary Fusion in vivo high throughput cloning technology (patent pending) is an extremely easy PCR cloning method:
- Simply amplify your gene of interest by PCR using a primer pair flanked with short arms homologous to the expression vector.
- Next, add 1µl of your PCR product to the engineered, ready-to-use Cloning Cells and
- Immediately proceed to transform the cells.
- You do not need to buy the expression vectors because the expression-optimized E Coli or mammalian expression vector has already been built into the Cloning Cells. (Note: to use your own vector, we can build it into our engineered cloning cells as a custom service).
For E Coli expression, the verified positive clone will be directly used for T7-based or nonT7-based protein expression (depending on the built-in vector), thus eliminating the second transformation into an expression strain, such as BL21.
Using our Fusion In Vivo technology, We successfully made a large microRNA expression clone collection for one of our clients with a very fast turnaround time and at much lower cost compared to other cloning technologies. We can do the same thing for you! If your collection can be PCRed from genomic DNA or other templates, we can build a collection of shRNA, miRNA or any other.
Advantages of Gentarget’s Fusion In Vivo, EcoTm Cloning Products and Services:
- It is the easiest and most cost-effective PCR cloning method available: simply add 1µl of PCR insert into the provided cells for transformation regardless of the insert’s size and concentration
- No need for the tedious bench work of preparing the vector backbone
- No need for any enzymes or tube reactions
- Precise directional cloning of PCR products without any extra protein sequences (except affinity tags)
- It does not matter if the PCR products have poly-A overhangs at their 3′-ends. The ploy-A overhang, if there is one, will be removed in the cloning step.
- Compatibility with a wide range different PCR sizes, from 200bp to 10kb
- Engineered E Coli and mammalian expression vectors that produce high protein yields
- Exceptionally adaptable for high throughput cloning