DNases and RNases are notoriously tough enzymes. :)
But, I think people worry more about these than they should, too. If you use deionised water, then hopefully there are not enough ions around to 'feed' the enzymes before the reaction is prepared and ready to go. Just remember to keep the tubes chilled/on ice before and after the reaction, and consider adding a _small_* amount of EDTA after the reaction to mop up any leftover buffer ions, if you're sure you're finished doing enzymatic things.
Remember though that people were doing molecular work long before the nature of DNases and RNases were even properly figured out. DNases in particular are rarely blamed for bad times, though I know people working with RNA who have blamed RNases for a lot.. but then, you never know. Maybe the real problem was something else and they just assumed RNases were the problem for cargo-culty reasons.
*EDTA is a strong chelator and can be a nasty pollutant if overused, so try to make an effort to use only the amount you need, if you use it at all. Ideally you'd familiarise yourself with molar quantities of whatever ions you're trying to clear up (e.g. Mg++, Ca) and you'd only add a proportional amount of EDTA to the buffer or even less.
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Sep 10, 2020, 14:00 by antoni.nicolauv@gmail.com:
About the mineral oil that you find, I would say that could be a problema the DNase, RNase and protease present in it.Check uct profile of the mineral oil PCR reagent: https://www.sigmaaldrich.com/content/dam/sigma-aldrich/docs/Sigma/Datasheet/2/m8662dat.pdfit says:Product ProfileDNase, RNase and protease: None detectedSuitable for use in the Polymerase Chain Reaction (PCR).But, maybe if you warm up the mineral oil that you found during 30 minuts at 100ºC the enzymes should denaturalize, don't they?El sábado, 5 de septiembre de 2020 a las 5:57:03 UTC+2, Tom De Medts escribió:Hi NicVied,Can you please share a link to this pocker PCR, thank you.Are you going to later mineral oil in lieu of a heated lid?If yes, then will you buy it from a scientific lab supplier or just from some marketplace like amazon?I found this link for the least expensive mineral oil - https://www.amazon.com/Mineral-Pure-Organic-Ingredients-Utensils/dp/B07G5KVW4G/ref=sr_1_33?dchild=1&keywords=mineral+oil+PCR&qid=1599277973&sr=8-33but I am not at all sure if this will serve the purpose...I look forward to your response.Cheers!TdMOn Fri, Sep 4, 2020 at 3:36 AM NicVied <antoni....@gmail.com> wrote:Thanks you all, finally I bought the pocket PCR. It's really cheap but without "hot lid", old school style. I will let you know how it work!El miércoles, 19 de agosto de 2020 a las 4:36:51 UTC+2, jfo...@udel.edu escribió:It depends to some extent on how "diy" you want to be and what your skill set is... I cobbled together a serviceable thermocycler that can do a half dozen epi tubes from the cooling block from an old HP workstation I had in my attic, a cartridge heater, an arduino w/ thermocouple, and some other electronic bits and pieces I had lying around. Material cost was basically nothing, but for the time I spent tinkering to make it serviceable I could have picked up a consulting gig and made enough money to buy a nice, shiny, new one...
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