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  • sequalprep normalization of amplicons for titanium

    Hi

    Has anyone used an invitrogen sequalprep normalization plate to normalize amplicons for multiplexing on 454 titanium? I noticed it was used by Kirk-Harris et al (Applied and Environmental Microbiology, 2010 76:3863-3868) in comparing normalization approaches for 16S amplicons.

    I'm intrigued because it's billed as one-step amplicon purification and normalization but it doesn't seem to include size exclusion/gel purification steps. I would be worried about small amplicons slipping through then dominating after subsequent amplification.

    Anyone tried it?

    Cheers,
    Paul

  • #2
    *disclaimer* I developed this product while at Invitrogen. This is neither an endorsement nor condemnation of this product. *disclaimer*

    There is not a major size selection performed with this purification technology. Primers will be removed, while larger primer dimers will not. Performs best with fairly clean PCRs.

    Nice to see that paper, I hadn't caught it when it came out. Thanks!

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    • #3
      You wouldn't happen to be aware of an alternative to this method would you? They can be a pain to get hold of, back order dates have slipped repeatedly (popularity?)

      I know the binding capacity of SPRI beads is in vast excess, but I wonder whether something homebrew which included size selection as well as normalisation could be concocted with them?

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      • #4
        we used these just-a-plate ones a few years back. They seem to be the same idea and worked fine as a I remember. http://www.charmbiotech.com/product.html

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        • #5
          Thanks Paul! That's an ironic retro GoDaddy style website

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