I found in a paper that they added dUTP and uracil nucleotide glycosylase to the PCR mix for better performance of the PCR.
Hi!
Sounds like the system to prevent contamination of PCR reactions by carry-over from previous reactions. Adding dUTP results in PCR products with U instead of T. If this product is carried over into a new PCR, it might outcompete the intended DNA template. The uracil nucleotide glycosylase will remove the Us in these carryover contaminations, preventing their use as template. Once the PCR starts, the enzyme is inactivated, so the new U-containing product can be amplified from the intended template.
Hi!
Sounds like the system to prevent contamination of PCR reactions by carry-over from previous reactions. Adding dUTP results in PCR products with U instead of T. If this product is carried over into a new PCR, it might outcompete the intended DNA template. The uracil nucleotide glycosylase will remove the Us in these carryover contaminations, preventing their use as template. Once the PCR starts, the enzyme is inactivated, so the new U-containing product can be amplified from the intended template.
Hi.
Thank you.If I added them to the PCR premix , could the quality of the band in the gel get better?
Hi!
I would not expect this, but I also cannot rule it out.
If you do not have PCR porducts containing uracil in your setup, the enzyme should not do anything.
The dUTP is another matter. I would not use that if ypu plan using the product for cloning, for obvious reasons. If you just want to use the PCR for tests, you can give the dUTP a try and see what happens.
Hi
In the protocol Iam using dUTP and uracil nucleotide glycosylase are being used. Iam using PCR as a diagnostic technique.
Then it is a good idea to use both, dUTP and the glycolase, as you will have less problems with contaminations from prior PCRs.
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