Storage Condition

Store at -30 ~ -15℃ and transport at ≤0℃ .

Components

Description

AbTaq DNA Polymerase is the monoclonal antibody modified hotstart Taq DNA Polymerase. The antibody can effectively block the activity of DNA polymerase below 55℃ , and irreversible dissociation occurs at high temperature to restore the activity of polymerase, thus effectively avoiding non-specific amplification at low temperature. This product is mainly used for qPCR, especially Taqman probe method.

Definition of activity

One unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid-insoluble material in 30 minutes at 74°C.

Quality Control

Endonuclease Activity

The product was tested in a reaction containing a supercoiled plasmid DNA substrate. After incubation for 4 hours at 37℃, there was no significant change of the DNA substrate by agarose gel electrophoresis.

Exonuclease Activity

The product was tested in a reaction containing a DNA substrate. After incubation for 16 hours at 37℃ , there was no significant change of the DNA substrate by agarose gel electrophoresis.

Residual Host DNA

The product was tested by TaqMan qPCR with primers specific for the E.coli 16S rDNA , and the results show that the E.coli genome residues less than 10 copies.

Method of application

1.Taqman qPCR reaction system:

a.The recommended final concentration of primers is 0.2 μM, which can be adjusted at 0.1~1 μM when the effect is not good. Primer length should be set at 18~25 bp and GC content at 40%~60%. The optimal amplified target fragment is generally 80~200 bp, which should be designed to avoid hairpin structure, dimer and other complex structures, and should be as far as possible across the intron region.

b.The final concentration of the probe is recommended to be 0.25 μM. If the effect is not good, it can be adjusted at 0.1~1 μM.

c.The dosage of template should not exceed 10% of the total reaction system, and the recommended dosage of sample is 1~2 μl. Different types of DNA templates contain different number of target gene copies, and gradient dilution can be carried out if necessary to determine the optimal amount of DNA template addition.

2.Taqman qPCR Cycling Conditions: