[PStepper]

Hi everyone,

this is the main discussion space for BOMB protocol #6.1 TNA extraction from mammalian cells using GITC lysis.
Here you can post all your questions and feedback!

[PStepper]

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[Jstees]

So I see a volume of beads to use for the protocol, but I can’t seem to find a concentration.

Does anyone know a working concentration? Or better yet, the concentration used to optimize this protocol?

[Tomek]

Dear Jstees,

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 volume of water (this is our stock stock). From this (or further dilutions in water) we take different amounts for purifications to benchmark the beads. And basically, using this information we take the appropriate concentration of beads for the preps.

We state volume in the protocol, simply because the concentration of the buffers/solutions was optimized (and should not vary too much for preps to work nicely).

Hope this helps,

T.

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