Does anyone do plasmid minipreps on their liquid handler, and if so might be willing to share details of what works for you?
We have an OT-2, rather than large deck liquid handlers like a Tecan or Hamilton, so Protocols that involve the OT-2 would be especially appreciated!
I’ve heard the MPE2 pressure module from Hamilton is really good for automated minipreps.
I think the problem with the OT2 in this case is you generally need to move product plates between locations whether you’re doing column or magbead protocols, and the OT2 doesn’t have an integrated gripper.
Or you could try something like what @JDesRo designed
Thanks! Forgot to mention: Our OT-2 is integrated with an external robotic arm. So can move plates around onto equipment that is external to the OT-2
Curious but have you ever used the Biotage Miniprep Columns?
I wonder if they are developing something for the OT-2 or if they already have compatible tips.
I’ve heard of such tips, but never tried them myself or on our OT-2.
I’m expecting they’re silica based tips. Was brainstorming that an E. coli lysis with non-ionic detergent + lysozyme + an anion exchange resin, would be the way to separate the plasmid from genomic DNA, without needing a flocculation / centrifugation / filtration step like typical chaotropic salt + silica plasmid minipreps go.
Maybe if they have anion exchange based tips, could work.
Edit: They do have such tips, but the explicitly Opentrons compatible ones only have 20 µL resin, which is probably not very usable for plasmid purification:
They do sell 1 mL tips that have ~300 µL resin, but unclear if those fit on the OT-2
They have a white paper where they mention a method involving E. coli on a Hamilton. No centrifugation needed.
Link? Would be happy to read up on it!
Might not be able to say, but what robotic arm do you use? I’m working with 2 OT-2’s and the lack of movement is a hinderance I’m looking to get past!
We’re using a Peak Robotics KX-2 robot arm. Long story short, we mounted the arm to custom 80/20 tables that have Trespa tops, and are developing a SiLA2 driver to control the arm (currently we just control it via a Python script, which could be improved)
fyi I think the new OT-3 is supposed to have a gripper (see here), but I know that’s not helpful now.
Ahh cool! Thanks for the info will have a look, appreciate it.
Do you know when that’s supposed to be formally announced? I also saw 96 head in some of the early firmware
Omega biotek makes a magbead based plasmid miniprep kit that I’ve had success with on a Biomek. I’ve seen Omega biotek protocols on the opentron protocol library before but don’t know about the miniprep kit specifically, it should be possible though, so long as you have an attached centrifuge and either a magplate or the opentrons magnetic module.
I’ve also heard the positive pressure system works well with columns but haven’t tried it myself. In my hands centrifugation and vacuum approaches were pretty unimpressive when paired with silica column-plates. They seemed to lose a large volume of eluent compared to normal spin columns, which resulted in A) loss of valuable plasmid DNA and B) lower concentrations of DNA
PS I think using certain kits (Galenvs) it would actually be possible to do a no-centrifuge miniprep with just the standard opentron magnetic module and no robotic arm
When I was at SLAS and talked to Opentrons, I was told to expect a lot more news “This summer” about the OT3, but they wouldn’t get any more specific than that.
96 well blocks from thermo and vspin centrifuge.
Should be an app note from cybio on the felix about this.
Omega has some kits standardized on the Microlab PREP, which is in the same general space as an OT-2 (we do have CORE grippers that can move plates between mag blocks and the heater/shaker that come included with the system)
Zymo has purchased a couple PREPs and is developing protocols for their kits. No idea if they are planning on publicly releasing those ala Omega, or if it just something they’ll have in their back pocket for customers who ask them for it.
A couple of other kit vendors are talking with us about buying some PREPs to do the same thing.
I’m going to give @cyancey a plug here and say you should hit him/Automata up in this case–they do some SCARA arm integrations and have a cool “automation lab bench”
we tested them and their final material quality is not that great
WE are trying to do the same on Hamilton. If anyone has any success, will definately love to hear. we tried several magnetic beads and there final quality is not that good compare to columns. we are still looking in it. Happy to discsuss more