Is it necessary to DEPC treat ? - (Nov/29/2006 )
Is it necessary to DEPC treat and autoclave the tips microfuge & PCR tubes if you are opening it from a new bag wearing gloves when the pack clearly says the material is DNase & RNase free?? My concern is I have been using 0.1% DEPC which looks like interfears with my PCR reactions!
No, as long as the bags or packages have no holes...and remember to reseal the packs!
never DEPC treate anything to do with PCR. i only do that when i'm running northerns or a nuclear run on.
i do use filter tips (ART) for using PCR though, and they're pre-sterilised. my pipettes are washed over in bleach too.
may the god of PCR smile on you.
if it is written that it is DNAse and RNAse free on the packet of ur tip and tube then no need to treat them with DEPC. just transfer the tip and tube with a RNAse free forcep in a tip box and tube box respectively and then just autoclave and dry them induvidually. so that no other of ur labmate can touch the box of tip and tube.
and one more thing, if u treat with ur tip and tube with DEPC and once u autoclave it , i think there is no DEPC , it reacts with RNAse.......and inactivate RNAse. so what u make worry.
well i dont know very much about this...........if someone can give better explanation or suggestion it could be help.
i only autoclave normal tips and all PCR stuff. keep them in PCR place only and reseal, as minnie said. no problems here.
back in lebanon duing my masters I was told to never use anything for PCR unless it is DEPC treated. here noone DEPC treat so neither do I
well it's because atucoalving DEPC generates alcohol which may interfers with PCR.
Why use DEPC if you are not using RNA in your experiment?
If you use DEPC at all, it is prudent to autoclave solutions and heat-resistant objects after DEPC treatment. Autoclaving decomposes the DEPC (use loose caps on bottles to allow CO2 to escape). DEPC messes up bases of nucleic acids (especially adenosine), ruining their ability to base-pair.
Henderson R.E.I, Kirkegaard L.H., &Leonard N.J. 1973. Reaction of diethylpyrocarbonate with nucleic acid components: adenosine-containing nucleotides and dinucleoside phosphates. Biochim. et Biophys. Acta, 294, 356-364.