- This topic has 5 replies, 3 voices, and was last updated March 11, 2021 at 2:30 am by .
- Posts
Hi everyone,
this is the main discussion space for BOMB protocol #4.3 BOMB Gel extraction
Here you can post all your questions and feedback!Hey Everyone,
Our lab is really enjoying the beads, they work great. We have come across some issues when using them for gel extraction however. When we add isopropanol to the dissolved gel and the carboxylated beads we use, the beads tend to clump together. No vortexing, pipetting or even mechanical disruption tends to break the beads apart again at this point. The yield is next to zero because of this. Does anyone have experience with these issues?
Thanks,
Timon
Admittedly, I have encountered this issue myself. Exactly as you describe.
At the time I did not find a solution right away and resorted to doing a PEG size cleanup instead.I will ask my colleague who contributed this particular protocol whether she has any idea as to how this can be solved.
Could you tell me the % of your gel and the weight of gel you dissolved per sample. Assuming that the beads unwantedly bind some of the agarose in the Isoprop sol. it might be that there is simply too much agarose relative the beads.
Cheers
Tim M
Hello Tim, thanks for you response,
I usually use 0.8% agarose gels, and try to dissolve 300mg of gel as a maximum for gel extraction. We tried to double the length of the gel dissolving step, but that did not prevent it. Neither did washing the carboxyl beads in advance, or heating the beadfilm in an attempt to dissolve remaining agarose. Hopefully your colleague has a solution!
Cheers,
Timon
- You must be logged in to reply to this topic.