We need to purify many oligos per day, however to buy a commercial automated oligo purification system is not an option since they are quite expensive, however we have a way to design and build one by our selves.
We know the best is to use columns and a manifold system coupled to a pipette dispensor. Has anyone built an in house device for automated or semiautomated oligo deprotection and / or purification?. What kind of vacuum manifold shall we use? Any specific directions or recommendations?
We use diaphragm pump with teflon diaphragm. Our device consist of two plates: upper plate with RP-columns for purification and lower plate which is installed after deblocking DMTr-groups for collection oligonucleotides.
For exhample see pictures on link below:
https://www.google.com/search?q=techniques+for+purification+of+oligonucleotides&newwindow=1&rlz=1C1CHRV_ruRU571RU571&espv=210&es_sm=93&source=lnms&tbm=isch&sa=X&ei=gGAZU_3zOtHV4ATfoYHYBw&ved=0CAkQ_AUoAQ&biw=1366&bih=630#facrc=_&imgdii=_&imgrc=8fpvSdUlncDXVM%253A%3BbhW6ITMOLiLIGM%3Bhttp%253A%252F%252Fwww.promega.com%252F~%252Fmedia%252Fimages%252Fresources%252Ffigures%252F6900-6999%252F6947ca_500px.jpg%253Fla%253Den%3Bhttp%253A%252F%252Fwww.promega.com%252Fproducts%252Fdna-and-rna-purification%252Fplasmid-purification%252Fwizard-sv-96-and-sv-9600-plasmid-dna-purification-systems%252F%3B500%3B611
We´ve used cartridges for oligo purification, but its not cost effective, Im thinking more in desalting using sephadex but in an autmated fashion, perhaps a robot. Any ideas?
Are your oligos in a 96 well rack format? How many oligos are you trying to Desalt at one time? You can use a vacuum manifold to Desalt. If you want to perform OPC, Desalt and Elute in an automated way, you can look at a robot but they are really expensive. There is an instrument called the Dr. Oligo Processor that automates all of those steps. The same company also sells a vacuum manifold that I mentioned earlier.
How are you performing your cleavage and de-protection? Are you using gas phase deprotection using ammonia hydroxide or Methylamine?
Hey, guys... I'd like to bring this topic back to life...
Any advances in sutomated purification systems for desalting and/or cartridges since 2014?
I've used a K&A P8... it's quite robust and easy to use and to perform maintenance... But is not that accurate, and also is only semi automatic, for you have to put the oligos once the cartridges are ready for the purification. Yields good oligos, neat and clean, though.
I've also heard about a Dr. Oligo Processor, saw a video, but never used. Seems to be more complete.
Any tips on other machines?
And mainly... any recipes for home made machines? It seems to be relatively easy, with valves, pumps, pressure... Dmitry E Zaytsev maybe you could provide more detail on the machine you made?
As I recall you can use an SPE vacuum manifold to desalt a sample (using Sephadex G15 cartridges, if they make them anymore)
Bruce Neagle you're right, we do use vacuum manifold... but i'd like an automated way to both dispense and drain reagents!
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