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  • Aliquoting AMPure XP magnetic beads recomendations

    I'm relatively new at working with RNA and I'm getting ready to make my first Library next week. Since I'm new to most these techniques and I want to be able to throw out anything that I feel might have been accidentally contaminated my standard operating procedure has been to make smaller aliqouts of all my reagents so I don't have to throw everything away if I make a mistake.

    I was just considering aliqouting the AMPure XP magnetic beads for this reason but I have never worked with magnetic beads before. Other than vortexing the beads to make sure they are evenly suspended before aliqouting are there any considerations I need to be aware of? For example, I'm assuming it is OK to store them in standard RNA/DNA low binding tube eppendorf tubes?

    I know this probably seems like a dumb question but the lab I'm in doesn't really have any molecular bio expertise and I'm learning everything from the product manuals, so any help would be greatly appreciated.

  • #2
    No need to even store them in low-binding tubes, regular RNAse/DNAse-free microcentrifuge tubes are fine. Also note that typically by the time you use the beads for library prep your RNA has already been converted to double-stranded DNA, so there is no need for RNAse-free "paranoia."

    Ampure XP beads can actually be used for RNA purification/concentration too, but that is probably a topic for another thread.

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    • #3
      just equilibrate the beads at RT for a while before mixing them (30 mins are enough for a 60 ml bottle), they contain PEG which is very viscous at 4 degrees. Mix really well and then make the aliquots. The binding capacity of the beads is so high that small pipetting errors or different amounts of beads in different aliquots won´t make any difference.
      A step further: save (A LOT of) money and make your own beads. Several protocols are available, like the one described in PMID: 22267522.

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