I had to switch to my PC I didn't see an option to attach an image from my phone.
Would you recommend I raise or lower the annealing temp?
I can try some variations on elution volume, but I don't have a way to measure plasmid density in my elution. I have done transformations successfully with good efficiency with my minipreps. Yesterday I took the remainder of the miniprep and added gelgreen to it then compared it to elution buffer without plasmid but with an equal concentration of gelgreen. The plasmid elution has clear florescence. I feel comfortable that my mini preps are working.
This is how the gels typically look, that's a 1Kb ladder in the left most well.
On Tuesday, March 1, 2016 at 9:17:38 PM UTC-6, Dakota wrote:
Your dilution looks fine.Less than 100bp is usually primer dimers. Check your annealing temp, make sure you have enough template. As Tom said you do have a pretty big amplicon, so it might take some fine tuning.Could you post a picture of your gels?You know your miniprep worked?On Tue, Mar 1, 2016 at 9:55 PM, David Ishee <midgard...@gmail.com> wrote:I don't know, it's less that 100 bp dimers are possible. My target and product would be much bigger.
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