Re: [DIYbio] Re: restriction digest confirmation

On Tue, Jul 16, 2013 at 12:37 PM, Nathan McCorkle <nmz787@gmail.com> wrote:
> So just do the math to bump the fragment up to the minimum detection
> concentration by adding more template... or after digestion, ligate on
> adapters (primers) to bump up the mass of ANY/ALL fragments. In the
> latter case, you'd then need to differentiate 6bp.

Actually, now that I think of it, as long as you use ~1:1 adapter to
template (concentration of all molecules in the digestion, or the
initial template multiplied by 2 assuming there are only two cut sites
associated with your double digestion) and incubate with ligase a
while, you shouldn't see any residual adapters.


OR maybe just assume the double digestion worked, ligate back to
circular, then PCR with a primer that ends with the 6bp site. If it
was deleted, you should get no PCR product.

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