Great to hear that you could adopt the protocols. BTW, you don’t need to synthesize the beads in anaerobic conditions. The synthesis protocol works well without argon or N2 environment.

It would be great if we could include midiprep or other protocols.

Cheers,

T.

Thank you for the question. The protocol 1.1 magnetic core particles are not stable long term. They have to be coated either with silica (protocols 2.1) or carboxyl (protocol 3.1) for full functionality. I guess if you were using COOH beads before, then you could use the #3.1 beads for coupling. Another chemistry would have to be…Read more

Sorry to hear that it still didn’t work for you. One thing that comes to my mind is that the iron salts could be old. Buying fresh good quality chemicals solved a similar issue for another user. And then the synthesis worked like a charm. Not sure if this is the trouble here, but small amounts of fresh salts one can buy relatively…Read more

Thank you for your question. Initially, we were hoping that people will manufacture the beads themselves and use for their research. However, we had so many requests for people to buy these, that we decided to offer both the beads and the racks commercially in the near future.

Best regards,

Tomasz

Thank you for your question. Indeed the concentration of the beads is not stated. We typically check the efficiency of the synthesized magnetic beads (titer the bead amount in the purification procedure and see the outcome), before we apply them for extraction from “real” samples.

After the synthesis, we mix the wet beads with same…Read more

Thank you for your message. Frankly speaking, I did not consider the possibility that TEOS might not be accessible to some, but you are right.

We did not try to use sodium silicates for coating, but it seems it is not too difficult to do, so we might give it a try sometime. If you are planning to try this before we do, it would…Read more

Sorry for the late reply. Thanks, it is in part reverse and forward engineering, but our hope, in the end, is that we made it accessible to everyone who would like to do cheap and efficient nucleic acid extractions using magnetic beads.

Regarding your question, there are quite a few possibilities to conjugate additional molecules to the…Read more

Thank you for your question. We did not try other coats than silica and carboxyl (MAA). However, instead of methacrylic acid you should be able to coat the core particles with hydroxyethyl methacrylate (HEMA) in the same kind of polymerisation reaction (simply to replace MAA with HEMA. This should provide a surface with OH groups.

No…Read more

To answer your questions,

1) Yes, from my experience if the ratio is not optimal, one gets precipitate which is also magnetic (not as strong thou) which then in washing gives brownish “color”

2) The solution should be clear and not cloudy. Not sure what exactly the “precipitate” suspension is, but removing them with filtration helps…Read more

Sorry to hear about your troubles with synthesis. We also occasionally had troubles like you are experiencing. There are a few things that are important here.

1) The correct ratio of Fe2+ to Fe3+ (otherwise proper MNPs have no chance to grow properly).

2) It can also be that the chemicals (iron salts) are old. The Fe2/3 solution…Read more

Sorry to hear about your troubles with synthesis. We also occasionally had troubles like you are experiencing. There are a few things that are important here.

1) The correct ratio of Fe2+ to Fe3+ (otherwise proper MNPs have no chance to grow properly).

2) It can also be that the chemicals (iron salts) are old. The Fe2/3 solution…Read more

There is no need for N2 or Argon! You should degas and heat up the solutions to ~80C and the synthesis should work. Degassing and the heat gets rid of a large part of O2 in the solution and does not disturb the synthesis too much.

Best regards,

Tomek

We never tried extracting NA from insects. I suppose this should not be too difficult to optimize. Probably you could follow the “Tissue extraction” protocols, where you would release the NA with ProtK treatment and then GITC buffer for cell lysis and NA capture.

Cheers,

Tomek

Not sure if the polarity matters that much. Would need to check. Nevertheless, in our magnetic stands all of them are aligned and have the same pole up.

Tomek

Tomek

Cheers,

Tomek

Well, we encountered a trouble like this when there was too much oxygen in the alkali solution or the iron II/III mixture was not well mixed. What I can recommend is to prepare the iron solution then mix it and filter with a 0.2 or 0.45 filter. The alkali solution we degas and heat up to ~80C which took care of the problem.

In…Read more

Best regards,

Tomek

Best regards,

Tomek

With the protocol you can get the RNA from TRIZOL solubilised samples directly without adding chloroform. Yet, if you added chloroform already and have separated the water and phenolic phases, you could add the RNA binding buffer to the water phase (where the RNA is), with 4 times the volume of the binding buffer to 1 volume of the…Read more

After synthesis (coating with silica of MNPs) and washing the beads, we resuspend them in 1:1 (volume) with water. After decanting the beads, we measure (more, less) what is the volume occupied by the beads and add that amount of ddH2O to them to resupend. This is our storing stock. Then, dependent on the application, we use different…Read more

Right. Thank you for spotting the typo, will correct it in the new version (indeed it should be 25%). Whether 7M NH3 in MeOH can be used instead, I don’t know. In principle the MNPs are already formed in the NaOH solution, thou I don’t know what effect the extra MeOH could have on the final reaction. In principle one could test it (or…Read more

No need to adjust pH. The Alkali solution will take care of that. The iron solution is also kept acidic (with HCl), to avoid iron oxide precipitation. We do degas the solutions using a vacuum pump and heat up the alkali to ~80C. Initially we were flushing the reaction bottle (and the solution) with N2, yet if it is degassed and preheated,…Read more

Thanks for pointing out the publication. Indeed it might be a neat way to make the core particles (I might give it a try soon). What I noticed in the publication is that the MNPs are relatively large (~50-100 nm) which is bigger than what we get with the coprecipitation protocol. In principle, the co-precipitation is also quite quick…Read more

Congrats. You posted the first non-BOMB-team member to discover the site.

Regarding the glassware, indeed we have used the flasks and funnels as Phil described. Yet, I have made the magnetic beads in all kinds of vessels before we had found our chemical glassware stock. Virtually any flask or bottle can be used for that. The key point…Read more