I did some calculations myself using some of yours (I had to go back and track everything I did) I found I had around 3.5 pmols per uL. (if everything went correctly)
So, for the gibson assembly, I had 2 different reactions. One was with about 10 pmol and the other was with about .5 pmol, of each fragment.
Now here comes the tricky part. I need to do a transformation today and I plan on using this protocol
https://github.com/cathalgarvey/biohacking-protocols/blob/master/E.coli%20Transformation%20With%20PEG%2BMgSO4.md
How much of the DNA should I use? I need to save some for verification, and some I need to use for transforming the Halobacteria! It is currently in a 20µl solution, so what do you guys think I should I dilute it to? Thanks for the help!
On Wednesday, March 13, 2013 8:48:45 AM UTC-7, Koeng wrote:
Hello all!--
I am using the gibson assembly and it requires a certain amount of dna, but I don't know how much dna my PCR reaction will create! Is there any equations, or recommendations on how much I should use if in my reaction I used .2 µM of primer in each reaction?
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