I was doing bacterial transformation of plasmid DNA. While adding LB media to the transformed bacteria, i accidentally also added around 400ul LB media to the pure plasmid DNA. The plasmid DNA was eluted from a filter paper and had 20ng/ul concentration, and its volume was approximately 24ul. I initially thought to centrifuge and pellet down the DNA but i was afraid if yeast extract in LB media would also come down. Then i thought of running it through mini prep column, but again DNA might also pass through column as LB media had water in it as well.
Can anyone please suggest how can i separate plasmid DNA from LB media? (The LB media had NaCl, tryptone, and yeast extract. It had no antibiotic. ).
you can try to precipitate it out through ethanol precipitation
protocol attached here:
https://projects.iq.harvard.edu/files/hlalab/files/ethanol-precipitation-of-rna_hla.pdf
The downside of this approach is that you might getting salt carry over, which could potentially inhibit enzymatic reactions downstream.
If it is a plasmid can be mini prep'd, I wouldn't bother to do this. Just grow another vial up and extract plasmid from scratch.
Good luck!
Either ethanol precipitation for a quick method or for a more precise method, do a transformation using a antibiotic selection and then miniprep DNA isolation.
You can just transform that plasmid again , and hopefully by using appropraite antibiotic selection you can repurify by using Miniprep.Good luck
Try grabbing it with a plasmid isolation kit and preserve the flow through, just in case. But as it seems that you don't have much material in the first place and you might need more late, transform bacteria with an aliquot and do a mini or maxiprep.
- Badges
- Science topic