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  • AMPureXP for ssDNA

    Hi Friends

    Has anyone used AMPureXP for ssDNA purification? I need to get rid of smaller cDNAs (<100 ntd).

    Biochembug

  • #2
    Yes it works for ssDNA. It also works for RNA. And DNA/RNA hybrids.
    --------------
    Ethan

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    • #3
      What volume should I use to eliminate low molecular weight (<100 ntd) cDNA?

      Comment


      • #4
        What I do is I mix 1 volume of DNA or RNA with 1 volume of AMPure XP and 1 volume of 30% PEG800 1.25M NaCl. That gives me a cut off of about 100 bp. If you search the forum there is a thread with more information on this with some guy that posted a nice experiment. Be warned that there is batch to batch variation with the AMPure XP beads.

        There are some protocols on my blog at http://ethanomics.wordpress.com/ which perhaps describe what I do a little better.
        --------------
        Ethan

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        • #5
          Thanks for the info Ethan. Will give a shot!

          Biochembug

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          • #6
            Originally posted by ETHANol View Post
            Yes it works for ssDNA. It also works for RNA. And DNA/RNA hybrids.
            Hi Ethan,
            Can you comment on what the cutoff seems to be for dsDNA vs ssDNA? Does it appear to be a function of molecular weight, strand length, or something else?
            Phillip

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            • #7
              There's a recent post on James Hadfield's blog detailing how the beads work and how the concentration of PEG/NaCl effects the size selection.

              Someone recently asked me, “how do SPRI beads work” and I realized I was not completely sure so I went to find out. My lab uses kits. Lots...

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              • #8
                Hi all,

                Great to see that you guys are keeping this thread alive. As pointed out by Phillip, I am looking for the cut-off of the single stranded DNA molecules on AMPure beads. Comments are always welcome!!!

                Thanks TonyBrooks for that like.

                Biochembug

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