FAQ

Q1. I could not see any band. What shall I do?

A1. You may have problem of Taq DNA polymerase or primer design. In most cases, the problem of the kit is occurred when primer design is improper or the quality of the template DNA or primer oligos are not good. If the problem is the primer design, please refer to our manual, pages 17 and 18 in which we identify the parameters for your proper primer design. Also, we suggest that you slightly change the annealing temperature or extension period in PCR.

Q2. Which DNA Taq polymerase do you recommend?

A2. This kit is DNA Taq polymerase sensitive. We strongly recommend DNA Taq polymerase from Roche (Cat. No. BM1418432) or Invitrogen (Cat. No. 10342).

Q3. How many target specific primers do we have to design for this kit?

A3. Basically we recommend the second nested amplification to obtain the best results. In order to do that, the user has to design three target specific primers: one for the first PCR, the second for the first nested PCR, and the third for the second nested PCR.

Q4. Is 2nd nested PCR necessary?

A4. If you obtained a specific band after the first nested PCR, we do not need to go to the 2nd nested PCR. However, if the first nested PCR did not generate a single distinct band, our protocol strongly suggests performing 2nd nested PCR.