Kun:LabNotes/ExonomeSeq/2007-12-30

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Exp. 1 Blunt-end cloning of the size selected PCR products from 12-23-07[edit]

Blunting of the DNA fragment[edit]

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

Blunt-end ligation[edit]

    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.