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Cloning:

4/13/14:

Done today: L-R reactions.

Protocol:

  • 1 μL of everything (μ = 03bc + alt + x in Word)

Conc:

Everything = 5 fmol  (5 fmol per μL)

Destination vector = 10 fmol

  • Put 1 μL of everything into the mix. Make sure all liquids coalesce
  • Then add 1μL of Clonase
  • After everything is added, pipet up and down. Careful not to add air bubbles. Then centrifuge.  If air bubbles form: tap or manual centrifuge
  • Incubate overnight at room temperature

Reaction tube numbers:

  1. (Kyle) TRE-LacOid_mKate in DEST 1-2
  2. (Erik) Hef1a-lacO_ NLS-eYFP in DEST 1-2
  3. (Erik) Hef1a-lacO_ eBFP in DEST 1-2
  4. (Shinjini) CMV_mKATE in DEST 1-2
  5. (Shinjini) CMV_eYFP in DEST 1-2

 

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