Standard method to define the unit concentration of Taq polymerase is based on incorporation of radioisotope-labeled dNTP, acid precipitation,and filtration. Since my lab does not have that facilities,I want to know whether there is any other method to define the taq polymerase unit concentration?
You can always titrate you polymerase stock and compare it to a commercial source. You would probably need to do this by real-time PCR and look for the concentration of polymerase that produces the same amplification profile.
I did real-time pcr by preparing different con of taq polymerase and compared the ct values with commercial taq polymerase which has 5 units per micro liter. Taq polymerase which has the same ct value as commercial one, nearly has 5 units per micro liter. And also end pcr product intensity in agarose was also same as the commercial taq polymerase. Therefore,this method would be a simple method to define unit con of taq polymerase.
- Badges
- Science topic