Just pass the eluate through the column a few times if you want to use a smaller volume

You can elute several times with molecular biology-grade water. Measure and pool your tubes, concentrate in the speedvac until you obtain a little volume in µL

Hi Maryam Mz

I suggest you warm your elution solution (usually TE buffer) to ~40 degrees celsius. After adding your elution solution to the column, leave it at room temperature for about 2 minutes. Then centrifuge at high speed.

The elution volume depends on the starting material. I have obtained high yields in the range of 1-2mg of plasmid (eluted in 2ml) from a 250ml bacterial culture.

Good luck!

Pushkal Sinduvadi Ramesh,

Thank you so much for your reply.

I am using the GenElute™ HP Plasmid Maxiprep, and I will follow your suggestion to elute the plasmid DNA before adding 0.1 volumes of 3.0 M Sodium Acetate Buffer (pH 5.2) and 0.7 volumes of isopropanol to the recovered plasmid. I will mix well by inversion and centrifuge at 15,000 × g at 4°C for 30 minutes, then carefully decant the supernatant without disturbing the pellet.

I will gradually add the elution solution, measure the DNA concentration, and continue adding until I obtain a very low DNA concentration. After that, I will proceed to the next steps. Additionally, I may heat up the elution solution and measure the DNA or plasmid concentration again.

This time, I am considering using 500 mL of bacterial culture, whereas last time, I used 300 mL (merging both 1:1000 and 1:500 cultures).

Please let me know if you have any additional recommendations.