Kun:LabNotes/ExonomeSeq/2007-12-20

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Revision as of 22:00, 20 December 2007 by KunZhang (talk | contribs) (New page: ==Exp.1 Cloning of the size selected fragments from 12/19/07== ===Blunting of the DNA fragment=== 5X Phusion buffer: 1ul 10mM dNTP: 0.25ul DNA: 3...)
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Exp.1 Cloning of the size selected fragments from 12/19/07

Blunting of the DNA fragment

      5X Phusion buffer:   1ul
      10mM dNTP:        0.25ul
      DNA:              3.65ul
      Phusion:           0.1ul
      72C 10min

Blunt-end ligation

    pCR-Blunt             0.5ul
    Blunt PCR product       2ul
    10X Ligation buffer   0.5ul
    Sterile water         1.5ul
    T4 DNA ligase (4U/ul) 0.5ul
    16C 1h -> 65C 10min -> 4C hold.