I read from Bitesize Bio website that Michael K. Danquaha, and Gareth M. Fordea (2007) published a paper for plasmid DNA production. This PDM can give a higher yield than the other E. coli medium, high quality of plasmid DNA, and is cost-effective.
My questions are
1. Does anyone ever use PDM (Plasmid DNA Medium) for E.coli plasmid preparation? and can this medium give a high yield or not?
2. Can a plasmid that prepared from this medium that contains a lot of mineral salts interfere with the downstream process for example: restriction analysis or ligation reaction or not?
The formula of PDM:
0.79% Tryptone, 0.44% Yeast extract, 1.0% Glucose, 1.28% Disodium phosphate 7H2O, 0.3% Monopotassium phosphate, 0.024% Magnesium sulfate, 0.05% Ammonium chloride
Thank you,
Tatpong
P.S. I already attach the paper file with this mail.
Honestly unless you are making DNA commercially in huge batches, the standard LB, TB and YT media are just fine. You will get more than enough high quality DNA to do whatever you like...
PDM is a lot like standard LB, but watch out for the glucose in PDM, as it will acidify your culture if you add too much or grow it too long, and this will result in cell death ... but even this only effects chr DNA and plasmid DNA much less.
If you purify your DNA with a column based kit (like Qiagen or Thermo) you don't need to care about whats in the medium as it will be gone anyway. If you don't use a column, the final ethanol precipitation will get rid of all the residual mineral salts.
Dear Soltes,
Thank you for your suggestion.
Now, I already prepare PDM. I will try to use this new medium, but my advisor suggest like you that LB is fine, not necessary to use the other medium.
The health of your culture and the amount of lyzed biomass are critical factors to obtain a good amount of pure plasmid. And the choosen media impact these two parameters. We developed our oown media. My advice to you: forget about the DIY kit option! Send us your plasmids as we can prep them for less than it takes you to do it. We typically obtain better results than you would with a kit in terms of DNA quantity and quality. See for yourself at https://www.biozilla.bio/plasmid-dna-purification
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