Does anybody know what the composition of the elution buffer is? Shouldn't that be almost water?
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When using Ampure XP beads, the included protocol says to dry for <5 min after the EtOH wash to avoid beads drying out too much and cracking. However most online protocols I see, plus Illumina's Truseq protocol (which uses Ampure XP) all say to dry for 15+ min until the beads crack. Anyone have any experience what difference this makes?
Also most protocols say to use fresh 70% ethanol to wash, but the Truseq calls for 80%. My understanding is that the higher ethanol concentration might be less efficient at washing away smaller molecules. Has anyone played around with this?
thanks
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Originally posted by katsigner View PostCan the beads be reused?? Any idea?
Beckman Coulter can't be happy about this idea. At least they may argue that you need to decontaminate the beads before binding other samples.
The homebrewed buffer seems to be promising; maybe someone can do some tests.
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Originally posted by edawad View PostWhen using Ampure XP beads, the included protocol says to dry for <5 min after the EtOH wash to avoid beads drying out too much and cracking. However most online protocols I see, plus Illumina's Truseq protocol (which uses Ampure XP) all say to dry for 15+ min until the beads crack. Anyone have any experience what difference this makes?
Also most protocols say to use fresh 70% ethanol to wash, but the Truseq calls for 80%. My understanding is that the higher ethanol concentration might be less efficient at washing away smaller molecules. Has anyone played around with this?
thanks
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Originally posted by chronicle View PostHas anyone tried this out yet or had any thoughts? I'm getting ready to start using Illumina's Trueseq kit as well, and this struck me as a little odd too.
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Originally posted by Heisman View PostI've done a bunch of bead purifications. I see pretty good results either way in terms of drying time. One thing to keep in mind is that if you use 70 or 80% ethanol, the ethanol will evaporate first and the last liquid you see will be water. So it can be a tiny bit wet and you can feel pretty confident that you have gotten rid of all of the ethanol. No idea about which ethanol percent to use.
Woops - should have noted - do you use a short dry time (<5 min like the Ampure XP protocol calls for), or do you go for a longer drying time (i.e. 15 min) as specified in the Illumina protocol? It seems like for such a small volume, this is a fairly excessive dry time, but I'm not sure how it will affect the Ampure Beads.Last edited by chronicle; 10-07-2011, 04:21 PM.
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Originally posted by chronicle View PostAwesome! Thanks for the prompt reply! Hopefully I will have similarly good results.
Woops - should have noted - do you use a short dry time (<5 min like the Ampure XP protocol calls for), or do you go for a longer drying time (i.e. 15 min) as specified in the Illumina protocol? It seems like for such a small volume, this is a fairly excessive dry time, but I'm not sure how it will affect the Ampure Beads.
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In my case the drying time is making only resuspension time a little bit longer and usually to be really sure that it's dry I dry beads 30 min under vacuum and nothing is happening to the product. But I have to note that mostly I'm working with the PCR products so maybe there are more stable.
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Originally posted by Yevaud View PostIn my case the drying time is making only resuspension time a little bit longer and usually to be really sure that it's dry I dry beads 30 min under vacuum and nothing is happening to the product. But I have to note that mostly I'm working with the PCR products so maybe there are more stable.
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Hi,
This question is related to AMPure XP but comparing it with the RNAClean XP. In theory the have different features:
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but some people told me that they are the same but with different buffer. Any idea or experience on this issue.
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Originally posted by chronicle View PostHas anyone tried this out yet or had any thoughts? I'm getting ready to start using Illumina's Trueseq kit as well, and this struck me as a little odd too.- Gina
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