I had Phusion HF buffer, dNTPs, Q solution and DNA primer stocks (100uM) exposed to UV this morning. Unsure how long the UV was on for. Has anyone had any experience with whether they would be ok to use?
your primers will be fine. Yes they are DNA, but single-stranded. UV exposure affects double-stranded DNA, causing thymidine dimers, thus not a problem for primers. Your dNTPs will also be okay, so should your buffer. It is not unusual for some labs to UV treat these (really targeting their containers...) prior to use.
Usually DNA + UV = dead DNA...and the more the exposition time, harder the damage.
I think primers are the affected ones in this case.
Anyways, if you are not sure how much time was, just get a positive control DNA and try a PCR, maybe was not even enough exposition time.
Good luck!
I think you should not use these reagents. It affects DNA (thymine dimers), its a known mutagenic.
your primers will be fine. Yes they are DNA, but single-stranded. UV exposure affects double-stranded DNA, causing thymidine dimers, thus not a problem for primers. Your dNTPs will also be okay, so should your buffer. It is not unusual for some labs to UV treat these (really targeting their containers...) prior to use.
It is my preferred way of neutralizing any contaminating DNA. I routinely expose my PCR mix to 5 minutes UV-treatment on a trans-illuminator before adding the DNA sample and the polymerase.
in case of primers and other PCR reagents UV has no effect but when it comes to DNA definetly UV exposure affects double-stranded DNA, causing thymidine dimers.
"The rates of dimer formation and the steady-state levels at most dimers are similar in single- and double-stranded DNA." See ref....
http://www.jstor.org/stable/3575688
Pyrimidine dimers form between two adjacent stacked prymidines on the same strand. Though this may be more efficient in the context of dsDNA there is no reason I can think of why it cant happen to single stranded DNA....
http://www.nature.com/scitable/content/pyrimidine-dimers-result-from-ultraviolet-light-a-19185
My guess is some small fraction of the primers are messed up. The PCR will probably still work though. If I were you I would replace the primers anyway, better safe than sorry mutation-wise. But primers are cheap right?
In my frustration of always getting a contamination, I exposed the primers and buffer to UV light for 30 minutes before using them. The primers were still working after UV exposure. The sad part was I was still getting contamination.
Jasper John A. Obico
We had similar issue with contamination. UV-treated reagents were giving us smear on the gel. Try HPLC-purified primers.
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