Hi I am a honours student starting only this year. I have done PCR about 3 years ago for about 2 months. The reactions then had worked well. I had +1 reaction volume to all my master mix to reduce pippetting error and also to prevent running out of Master Mix for the last tube. I never had problems with the pippetting.
Now I have started new work on my honours project and has been having PCR failures for at least 10 reactions.
For the first few reactions, we thought it was pippetting error because I was new. Then after that I changed the primers. I designed and bought new ones. I have tried a few of them and the result is still the same. My supervisor disagrees to my +1 method and insist that I need to work on my pippetting technique. It was alright sometimes and more often than not, I don't end up with the correct volume of leftovers and sometimes run out of MM for the last tube.
Is pippetting error really detrimental to a PCR reaction or is it really just my primers?