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changes.mady.by.user Andrew Hirning

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changes.mady.by.user Sara Molinari

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  1. 10mM dGTP (can be any other dNTP)
  2. T4 ligation buffer
  3. NEBuffer 2.1 (NEBuffer 2 with BSA)1
  4. At least 18μl of spin column purified dsDNA
  5. BSA 2
  6. T4 DNA polymerase

Protocol

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  1. Though NEB says that T4 DNA polymerase retains full activity in CutSmart buffer, I believe this references the polymerase activity, not the exonuclease activity. SLIC did not work well with CutSmart buffer, but works very well with Buffer 2.1 -AJH
  2. NEBuffer 2.1 already contains BSA no additional one is needed