User:Karmella Haynes/Notebook/BioBrick cloning/2009/11/29

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11/29/09 ✓ Cultures: KAH99(2), 100(2), 101(2), 102(2), 103(2), 104(2) ✓ Oligo annealing: KAH012 (phosphorylate, anneal) Oligo annealing KAH012 ---> Phosphorylate oligos, then anneal overnight DNA oligos (100 μM) up to 21 μL (3 μL ea.) 10x PNK buffer (NEB) 3.0 ATP (10 mM) 2.0 T4 PNK (NEB) 2.0 dH2O 2.0   30 μL --> 37°C/ 1 hr. --> Add 4 μL 500 mM NaCl --> 100°C waterbath/ 10 min. --> Cool to R.T. overnight