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  • Clontech Low Input Library Prep Kit

    Hello,

    Anyone used Clontech Low Input Library Prep Kit (Cat. No. 634947)? It is suggested for the lib prep following cDNA synthesis/amplification with the SMARTer RNA-seq kit. But I think it can also be used for ChIP-seq or other lib prep which starts from small amount of DNA. The performance of this kit, according to the manual, is really impressive – it generates library that is enough for Illumina sequencing from 100 pg of dsDNA with only 14 cycles of PCR amplification, and all the reactions are done in a single tube. Is it possible? Does anyone has some clue how this kit works?

    Best,
    Li

  • #2
    I, too, would like to know how it works.

    It looks like the clontech product is a re-badged version of the Rubicon Genomics Thruplex kit ( http://rubicongenomics.com/products/thruplex/ ). The manuals are identical.
    The Thruplex kit is cheaper, though.

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    • #3
      Thanks luc! The information you provided is very useful, there is actually a chart on Rubicon Genomics website:

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      • #4
        Thanks, Slees!

        I had not seen this page.
        I have to say this protocol is a thing of beauty (and seeming simplicity).
        The patent is very interesting, too.
        The present invention concerns preparation of DNA molecules, such as a library, using a stem-loop oligonucleotide. In particular embodiments, the invention employs a single reaction mixture and conditions. In particular, at least part of the inverted palindrome is removed during the preparation of the molecules to facilitate amplification of the molecules. Thus, in specific embodiments, the DNA molecules are suitable for amplification and are not hindered by the presence of the palindrome.

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