I started a PCR process and recall I didn't include the Primers of the gene of interest in the cocktail preparation process. The pcr was at the initial denaturing step (@1:00), Will my reaction be successful if I stop, and include the primers in the pcr mixture? To restart he pcr process?
Yes I expect it to work. The primers were not active and the enzyme activity will not be effected by the short heating step and all reagents are very heat stable. I would add the primers and restart the cycling
Agree with Paul. Just try it. Afterwards hou can check the specificity and efficiency by usual agarose gel tests. should be ok.
Yes, it will be fine to do so. I have done it myself and it didn't affect my final PCR product. I used MegaFi DNA polymerase cat.no. G896 from abm, but I imagine you can use other brands too.
Just give it a try! If it didn't work you will just have to re-run it anyway. I had a power failure (for the area, not the machine) and restarted the cycle anyway after the power came on an hour later. It was fine.
As a note, if you are planning to do anything with your PCR product (cloning or sequencing) and not just run out a gel to see bands you may want to re-run it just in case of any minor errors due to repeated template heating/cooling
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