stability of Phusion and Pfu at room temperature
Posted 12 September 2011 - 01:00 AM
by a very unfortunate accident, my tube of Phusion and Pfu enzyme were left for about 5 days at room temperature.
I am wondering whether i can still use these enzymes.
Have anyone ever encountered the same problem? Does anyone have any idea about how long these 2 enzymes can be stable at room temperature?
Posted 12 September 2011 - 01:50 AM
According to NEB- Phusion is stable at RT for up to 2 months (see FAQ here: http://www.neb.com/n...tM0530.asp#1423)
As for pfu- doesn't look promising, it says here (http://www.prospecbi...olymerase_8_135) that it is stable for 5 days at 10C so I guess it depends on what room temperature was for you.....
You could always just test some to see, but I guess it depends how precious your samples are.
Posted 12 September 2011 - 01:58 AM
Actually we gave a try on a positive control, both enzymes worked but bands on the gel displayed lower intensities whereas the PCR ran for 35cycles (which usually yield enough DNA to show a high intensity band). That's why i was wondering whether someone experienced something similar.
With the phusion we do cloning so it is important that in theory 5 days at RT should not have affected the enzymatic activity, as for Pfu we mainly use it for mutagenesis and generating blunt-PCR product so i hope it will be enough for this.
Thanks for taking the time to reply.