Summary

In the PCR system, the general polymerase has terminal transfer activity, usually adding A to 3', and TA cloning method links the PCR fragment to a vector DNA with 3'-T prominence. Therefore, TA cloning does not need to use primers containing restriction enzyme sequence, nor does it need to add joints or flat-end treatment to the amplified products of PCR. TA cloning was performed directly by blue-white screening.

requirements for experimental materials

In order to ensure the success rate of TA cloning experiment, please provide materials as far as possible:

1.Unpurified PCR products and PCR electrophoresis images are convenient to confirm the success of PCR.

2.The primer sequence was amplified by PCR to facilitate the verification of sequencing results after TA cloning was completed.

3.Enzymes for PCR amplification. If the polymerase containing 3'5'exonuclease activity is used, please provide PCR primers. We will reapply the PCR products to ensure TA cloning.

Service Items

service description

1.In the experiment of TA cloning of PCR products, the greater the PCR fragments, the more difficult TA cloning is. At the same time, some customers require full-length sequencing, we will charge according to the actual cost of sequencing and walking primers.

2.In TA cloning experiment, if there are special requirements for the vector, please inform the customer service personnel before the experiment.

delivery of results

To provide a freeze-dried plasmid (about 4 ug), the corresponding sequencing results and puncture bacteria, pMD18-T vector picture and sequence, TA cloning experiment report.