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TruSeq or Nextera kit for library prep?

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  • TruSeq or Nextera kit for library prep?

    Hello,
    Has anyone compared the TruSeq and Nextera kits for making RNA-seq libraries for Illumina HiSeq?
    I am trying to make libraries from relatively small quantities of totalRNA from plant cells. I would like to know if someone has done this comparison.
    Many thanks!

  • #2
    Co-ask.

    Thanks.

    Comment


    • #3
      I want to know too

      Comment


      • #4
        Is there a Nextera RNAseq prep kit? I don't see it on the Illumina website.
        What do you mean by "relatively small quantities" of RNA.

        --
        Phillip

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        • #5
          Truseq on 100ng input

          Hi everyone,

          Illumina currently only sell a Nextera kit for DNA library preps.

          The Nextera enzyme will only work on double-stranded DNA, so if you are considering testing it for RNAseq, I would recommend making double stranded cDNA first. Perhaps also consider testing the Nextera XT kit, since this is designed to work with Amplicons and very low input.

          We have tested the Truseq RNA kit down to 100ng total RNA input (for RNA with a high RIN number, >8) and the data has been good. How low are you trying to go?

          S

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          • #6
            No Nextera kit per se but there is an application note from 2010 - Application of Nexteraâ„¢ technology to RNA-seq library preparation. Nature Methods 7 (2010)
            Attached Files

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            • #7
              Illumina's Epicentre division has an RNA-seq kit that you may have interest in. http://www.epibio.com/products/rna-s...reparation-kit

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              • #8
                what properties does the DNA polymerase have in the Truseq DNA sample prepare kit?

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                • #9
                  You all should view these links. Nextera works for RNAseq with some caveats, but it has been published:

                  http://epicentral.blogspot.com/2010/...r-rna-seq.html

                  http://www.nature.com/nmeth/journal/...meth.f.317.pdf

                  http://genome.cshlp.org/content/earl...3.111.full.pdf

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                  • #10
                    Oop.. I missed the post above with the nat.meth paper.

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                    • #11
                      Epicentre has a Nextera-"powered" RNA-seq Kit, called "Totalscript", good for 1-5 ng of high quality total RNA input.

                      http://www.epibio.com/applications/r...eq-kit?details

                      Not much available in the way of posters, publications or the like at this time, though.....

                      Olaf

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                      • #12
                        I'm also a little bit confused. Nextera kits looks great! However,since Nextera is much better than Truseq, why does Illumina still sell Truseq kits?

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                        • #13
                          We can't say it's "much better", problem is that the Transposomes can't get within 20-50 bases of the end if each target (it's the biology of the system) such that coverage depth drops way off on the ends.

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