Deactivating Phusion For Storage of PCR products

[Biosynthetique]

I am copying my laboratory's parts collection for a collaborator. The library is naked DNA. I was going to use phusion PCR to amplify the parts and then just send him the dirty pcr products, but im wondering if I should deactivate the phusion polymerase at least. The only issue is that phusion doesnt denature with heat.. should I denature with something else, use a different polymerase, or not even worry about denaturing?

[Nathan McCorkle]

phenol chloroform extraction should do it, an silica spin-column would
probably be pretty good too

> --
> -- You received this message because you are subscribed to the Google Groups
> DIYbio group. To post to this group, send email to diy...@googlegroups.com.
> To unsubscribe from this group, send email to
> diybio+un...@googlegroups.com. For more options, visit this group at
> https://groups.google.com/d/forum/diybio?hl=en
> Learn more at www.diybio.org
> ---
> You received this message because you are subscribed to the Google Groups
> "DIYbio" group.
> To unsubscribe from this group and stop receiving emails from it, send an
> email to diybio+un...@googlegroups.com.
> To post to this group, send email to diy...@googlegroups.com.
> Visit this group at http://groups.google.com/group/diybio.
> To view this discussion on the web visit
> https://groups.google.com/d/msgid/diybio/82c5d060-ef86-4b07-bc71-5076ba1fed79%40googlegroups.com.
> For more options, visit https://groups.google.com/d/optout.



--
-Nathan

[Mega [Andreas Stuermer]]

How about adding TE? It complexes metal ions, and polymerases need them to work.

[Mega [Andreas Stuermer]]

It would be a gamble though, maybe the exonuclease part doesn't need them

[Jeswin]

On Thu, Mar 19, 2015 at 1:47 AM, Nathan McCorkle <nmz...@gmail.com> wrote:

phenol chloroform extraction should do it, an silica spin-column would
probably be pretty good too

PCR cleanup with a spin column sounds good.

[BraveScience]

If you clean up your PCR product and avoid eluition you could send directly the spin filter or column to your colleague.

A colleague of mine received plasmids in this way to avoid lengthy bureaucratic procedures to see its dna delivered.

And no water no degradation.

[Biosynthetique]

Thanks everyone. I am trying to avoid having to use many hundreds of spin columns - im amplifying 384 well plates and I just want to store it long term without doing a crazy amount of cleanup. 

[Cathal (Phone)]

How about using a protease?

--
Sent from my Android device with K-9 Mail. Please excuse my brevity.

[Mega [Andreas Stuermer]]

If it is only storage, why don't you store it dry? Most proteins won't work then so your DNA is protected

[Mega [Andreas Stuermer]]

Ah it was mentioned above already. You could use spin columms or jzst dry it in the wells or on filter paper