Experiment Gallery
Daily Notes
I ran a mini-prep today to get more of my miraculin + eyGFPuv plasmid from some e. coli that has it. Mostly following the bioland miniprep 2 kit, BUT:
- I start with a couple loopfuls of bacteria right from the plate
- I found that I can use a syringe filter (0.22um) in place of the first centrifugation step (getting rid of the goop)
- For the wash step, I made an adapter: syringe -> luer-to-barb -> bit of tubing -> barb-to-luer. Into this I could press the bottom of the column, such that sucking the syringe pulled the wash through
- For the final elution step, I used my dremelfuge since I needed what came through, then with the remaining lysate I did another go on the column. I tried (unsuccessfully) sucking the final elution buffer through into a 1.5ml tube with a needle + syringe. Then after I gave up and centrifuged it, I discovered that with a bit of force the syringe luer outer diameter just fits into the top of the column, so I can push it through and collect in future (tested with water, looks like it will work and is great apart from a little foaming/splattering).
All that to say, I am now convinced that you can do a miniprep with this kit without needing a vacuum manifold or a centrifuge or anything - just a syringe, syringe filter (useful for lots of other things) and maybe some type of tube adapter for the sucking part (but also TBH I think blowing through makes that obsolete, just blow through all the column steps). Hooray!
Does this help anyone besides me? Not sure, I think most people doing minipreps have centrifuges haha. But I'm glad I don't need to set up the dremelfuge if the dremel has something else set up in it.