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Recycle Those DNA Extraction Columns

Posted in: DNA / RNA Manipulation and Analysis
Recycle Those DNA Extraction Columns

You know those ridiculously priced and throw-away DNA mini, midi and maxi-prep columns? Well the good news is that you can actually re-use them if you are reasonably careful at regenerating them, with this simple and cheap method described in detail by Nagadenahalli B. Siddappa in Biotechniques in 2007.

Apparently these columns can be reused up to 20 times… perhaps more a guesstimate than a real number, but hey, who’s complaining?

Essentially the columns are treated with 1M HCl overnight, rinsed extensively with dH2O and then re-equilibrated with buffer from the kit you use (e.g. Qiagen QBT). If you are worried about finishing the buffers in the kits, look in the back of the manual, there are often simple recipes about how to remake them.

The authors allay concerns about plasmid carryover contamination by assaying for plasmid using RT-PCR and transfection assays which all showed zero carryover between recycled uses of the columns. In fact they showed that the plasmid DNA had been chemically sheared into low molecular weight fragments and could not be used as a template.

They also stated that prolonged exposure to 1M HCl (1 month) did nothing to the binding capacity of the columns, so even if you forget about it, there is no need to stress.

Our lab has modified the protocol to include passing warm water through the treated columns when regenerating to ensure that all DNA and any other contaminants are removed. Also pre-warming the elution buffer to 42oC or thereabouts seems to increase the elution of any plasmid DNA bound to the column, although what extra compounds it releases is unknown but probably negligible.

It may be our imagination, but we have often gotten higher yields of plasmid DNA from a treated reused column, than a new one.

Go ahead, stretch those budgets further.

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  1. Krzysztof Treder on January 18, 2017 at 12:22 pm

    Maybe for regenerated Quiagen silica columns, buffers described by Boom et al. (1990) can be used instead of proprietary N3, PB and QG? They were designed to purify DNA/RNA using silica particles or diatom earth, so should work the same on silica in the columns. Just guessing.
    Cool protocol anyway.

  2. Kanomi Sasaki-Capela on July 14, 2016 at 12:11 am

    Hello Everyone,

    The Qiagen manual states that the max yield for a midi prep is 100ug. I just performed a midi and ended up with 183ug of DNA. Normally a high yield is a good thing, but 183ug is unusually high. Has anyone yielded more than the described 100ug for a plasmid midi prep? Or is it possible some kind of contamination gave me a higher O.D. than the actual amount?


    • deemah dabbagh on September 28, 2016 at 9:49 pm


      I have never used the midi kit, but I have used qiagen’s mini and maxi kits and have ended up with conentrations as high as 300 ug/ml with the mini and 500 ug/ml with the maxi both with good quality DNA.



      • deemah dabbagh on September 28, 2016 at 9:50 pm

        And the OD was in the correct range both of the times.

    • Ruchita Selot on December 8, 2016 at 1:28 pm

      Hi Kanomi,
      I have used the midi kit and gotten plasmid yield of 220ug from one column. However, it gave poor transfection result so obviously the forced elution is messing up with the DNA! Have to figure out another way of getting the high concentration of DNA without compromising on the quality…
      Ruchita, Bangalore

  3. Desanka Maksimovic on November 12, 2010 at 2:13 pm

    Hello everyone!

    I’m using Sigma’s Spectrum Total RNA Kit. For the binding columns, I have found the protocol for recycling, but I’m wondering what to do with the filtration columns. I have an impression that they can’t be reused. Can I do the RNA instraction without them?


  4. Christ Fotiadis on October 8, 2010 at 6:31 am

    Hello to all of you

    I want to ask a question about the re-used columns. Did anybody tried to digest the plasmid from that columns? I tried a lot of times to digest it, but it was unable to digest. When i use a fresh new column with the same plasmid and digest it all goes well. I use the re-used columns for pcr without any problem. Do you know something about it? Thank You and sorry about my english.

    • Prashant on March 19, 2019 at 7:53 pm

      Could you elaborate “unable to digest”?

  5. Liam on April 15, 2010 at 3:34 pm

    Hi Bill

    This depends on your column. I have only used the Qiagen columns, although the Macherey-Nagel ones are probably the same. These are gravity based, but I don’t see why you couldn’t use vacuum filtration (I use vacuum filtration to clean them). I think you would just need to test it on equal volumes of the same prep to see if it is suitable.

    Good luck

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