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DNA cloning
With different DNA cloning methods , we can finish your cloning efficiently in a short time whlie guaranting the quality at the same time. With the technology of In-Fusion, the cloning of PCR products to vectors is more convinient and sucessful. There are many TA cloning vectors now which can Fully meet the next step experiments

The DNA cloning of bacterium is the process of creating multiple copies of an isolated DNA fragment or fragments by in vitro or in vivo methods. After ligation of exogenous gene and the cloning vectors with enzymes, the recombiants were transformed to bacterium host cells, followed by screening the transformants with the target gene. By cluturing the transfromants and purification DNA from it , a large amount of DNA can be obtained.

Service content
TA cloning of PCR products
As is known to all, most of the PCR products amplified with Ex Taq or LA Taq contain “A” and can be cloned to vectors which have “T” ends

The subcloning of target gene
The vectors which contain the target gene is digested with restrction enzymes and ligated with a commercial vector digested by the same enzyemes

PCR amplification of the target gene with plasmids as the template
One fragment of a vector is added one or two restriction sequences with PCR method. Sometimes it also means cloning different genes by PCR 

Multi- fragment efficient cloning
In-Fusion PCR Cloning Kits allow ligation-independent, directional cloning of PCR products into ANY vector, at ANY site of linearization. hese kits provide an easy, rapid, single-tube cloning process.


Vector construction
With different DNA cloning methods, we can finish your cloning efficiently in a short time while guarantying the quality at the same time. With the technology of In-Fusion, the cloning of PCR products to vectors is more convenient and successful. There are many TA cloning vectors now which can fully meet the next step experiments

The DNA cloning of bacterium is the process of creating multiple copies of an isolated DNA fragment or fragments by in vitro or in vivo methods. After ligation of exogenous gene and the cloning vectors with enzymes, the recombinants were transformed to bacterium host cells, followed by screening the transformants with the target gene. By culturing the transfromants and purification DNA from it, a large amount of DNA can be obtained.

The protocol for vector construction
1.Restriction enzyme digests:
Digest the expression vector or cloning vectors with restriction enzymes. The vectors which contain your target DNA or cDNA fragment is also digested with the same enzymes.

2.Gel purification of DNA fragments on an agarose gel:
Load 2-5ul digestion product on the well of the agarose gel, after the electrophoresis, check whether there are your target fragments.

3.Isolation and purification of DNA fragments:
Use a gel purification kit to purify the DNA fragments.

4.Ligation:
Ligate the target fragment and the vector with T4 DNA ligase or some other similar enzymes.

5.Transformation:
Transform the ligation products to a host, such as Escherichia coli, Pichia pastoris, and so on.

6.The screening of positive clones:
Identify the positive clones which contain the target fragment with the method of PCR. Alternatively, extracting plasmid from the culture of potential clones and digesting it with restriction enzymes is also a good choice. But the best choice is to sequence part or all sequences of the recombinant.
 

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