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  • odile
    Member
    • Sep 2009
    • 30

    #31
    Hi, thanks for your response. The problem is that I need to clean up the extract before I attach the adaptors (after the blunt end and adenylation steps). Do you know any other method that would retain the fragments <70bp?
    I was wondering if maybe people who work with RNA would have an idea?

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    • JUdw
      Junior Member
      • Dec 2009
      • 9

      #32
      Originally posted by odile View Post
      Hi, thanks for your response. The problem is that I need to clean up the extract before I attach the adaptors (after the blunt end and adenylation steps). Do you know any other method that would retain the fragments <70bp?
      End-repair, A-tailing and adpter ligation is combined in the Nextera sample prep protocol:

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      • odile
        Member
        • Sep 2009
        • 30

        #33
        I had writen to Nextera a few months ago because I wanted to use their kit without the shearing part and here is what they wrote back:
        "Unfortunately, 100 bp DNA fragments are too small for use with Nextera technology. The best products we could tell you to use would be the End-IT End repair Kit to generate blunt ends on the DNA fragments, followed by ligation of the adapters to the fragments with our Fast-Link DNA Ligase."

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