Could you label what's in each lane?
I've always found template DNA quality to have much more of an impact on PCR success rate vs annealing temp. I've had the same primers anneal from 48C all the way up to 60C fine.
A dirty miniprep and residual ethanol can affect PCR.
I've always found template DNA quality to have much more of an impact on PCR success rate vs annealing temp. I've had the same primers anneal from 48C all the way up to 60C fine.
If you have a mini-prep, you should have enough to run just your plasmid + your plasmid cut with restriction enzymes.
Run just your plasmid on a gel, and run your plasmid cut with a restriction digest.
On Tue, Mar 1, 2016 at 11:15 PM, David Ishee <midgardkennels@gmail.com> wrote:
One thing about my mini preps is I tend to still have some ethanol contamination in my elution. It's usually enough I can run a miniprep on a gel because it won't sink.
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