DyNAzyme EXT DNA Polymerase (1 U/µL) - FAQs

查看更多产品信息 DyNAzyme EXT DNA Polymerase (1 U/µL) - FAQs (F505L)

4 个常见问题解答

How important is the quality of my DNA template in long PCR?

Template preparation becomes particularly important when performing longer amplifications (>15 kb). Therefore, it is recommended to check the length of the DNA by agarose gel electrophoresis.

Does DMSO inhibit DyNAzyme DNA Polymerases?

10% DMSO will inhibit DyNAzyme DNA Polymerases to some extent. This inhibition may be overcome by using twice the amount of DyNAzyme DNA Polymerase.

What nucleotide analogues can I use with DyNAzyme and Phusion DNA Polymerases?

DyNAzyme II DNA Polymerase can use dUTP, biotinylated dNTPs, 7-deaza-dGTP, digoxigenin-dUTP, bromo-dUTP, radiolabeled dNTPs and ITP. DyNAzyme EXT DNA Polymerase and Phusion DNA Polymerase cannot read dUTP-derivatives or dITP in the template strand so the use of these analogues is not recommended. Use Phusion U Hot Start DNA Polymerase for amplification of dUTP and dITP containing templates.

Do DyNAzyme DNA Polymerases add the non-template dependent 3'-A overhang?

DyNAzyme DNA Polymerases are capable of adding a 3'-overhang. However, the efficiency at which the extra base is added is sequence-dependent. The extra base is usually A, however, the enzyme can also incorporate other bases or even more than a single base at a sequence-specific manner. To enhance TA-cloning it may be helpful to include an additional incubation period of 10-30 minutes at 72 degrees C to the end of cycle protocol.