Product description

Dual-Block Taq antibody V2 is a product developed by our company that targets and blocks both the 5’→3’ DNA polymerase activity and the 5’→3’ exonuclease activity of Taq DNA polymerase at two separate sites. During the temperature rise in the PCR pre-denaturation, Dual-Block Taq antibody V2 effectively prevents the leakage of Taq DNA polymerase activity and inhibits the 5’→3’ polymerase activity, thereby suppressing nonspecific amplification caused by primers under low temperature conditions. It also inhibits the 5’→3’ exonuclease activity, preventing the probes or primers from being cleaved and affecting the performance of the reagents. This product can fully release the activity of Taq DNA polymerase after heating at 95°C for only 30 seconds. Using this product to block Taq DNA polymerase significantly improves amplification specificity and the detection rate of low templates. No special inactivation treatment is required for the antibody when using this product, making it compatible with existing PCR reaction conditions.

Transportation and storage

Store at -25~-15℃ and transportation≤0℃.

Shelf life

2 years.

Quality control

• Purity: (SDS-page) ≥ 95%

• Endonuclease activity: Incubation of 2 μg of antibody with 1 μg λ DNA for 16 hours at 37°C resulted in no detectable degradation of the DNA as determined by gel electrophoresis.

• Exonuclease activity: Incubation of 2 μg of antibody with 1 μg λ-Hind III digest DNA for 16 hours at 37°C resulted in no detectable degradation of the DNA as determined by gel electrophoresis.

• Nickase activity: Incubation of 2 μg of antibody with 1 μg pBR322 for 16 hours at 37°C resulted in no detectable degradation of the DNA as determined by gel electrophoresis.

• RNase activity: Incubation of 2 μg of antibody with 1.6 μg MS2 RNA for 4 hours at 37°C resulted in no detectable degradation of the RNA as determined by gel electrophoresis.

• Mouse genome residue: No contamination of mouse genomic DNA is detected after 35 cycles of PCR.

Protocol

The concentration of this product is 3mg/mL. It is recommended to use 1 μL of antibody to block the activity of 15-75 U of Taq DNA polymerase. Mix and incubate at room temperature for 30 minutes (for volumes greater than 200 μL, mix for 2 hours, and customers should adjust the process for larger volumes). Store at -20℃ overnight before use.

Notes: The blocking ratio can be adjusted according to customer needs.

Precaution

• Avoid repeated freeze-thaw cycles and wear gloves during operation.

• Due to differences in the specific activity of different Taq DNA polymerase, the blocking ratio can be adjusted appropriately, with a blocking efficiency of over 95% being preferable.