User:Karmella Haynes/Notebook/BioBrick cloning/2010/03/15

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03/15/10 ✓ Assembly: KAH142 - DPRE+YFP reporter (KAH96) Assemblies KAH142: (1) DPRE/(E/S)/1740 + (2) KAH96/(E/X)/~6025 > Digests (Fermentas FD) Reagent Volume   30 μL/lane, 1% agarose DNA (plasmid) up to 25 μL 10x buffer 3.0 enzyme 1 1.0 enzyme 2 1.0 dH2O ---   30 μL --> 37°C/ ~30 min. > Measure conc.'s Sample OD260 260/280 ng/μL 1. DPRE (E/S) 0.863 1.93 43.2 2. KAH96 (E/X) 2.379 1.85 118.9 > Ligations Ligation Plate results (lig : neg crtl) mm/dd/yy 1. DPRE(E/S)/1740, 14 ng + KAH96(E/X)/6025, 25 ng KAH142 3:1 (Pick 6) 2. KAH96(E/X)/ 25 ng >100 colonies   1 2 Insert DNA 0.8 --- Vector DNA 0.2 0.2 2x lgn buf (Roche) 5.0 5.0 T4 ligase (NEB) 1.0 1.0 dH2O 3.0 3.8   10 μL 10 μL --> R.T./ 10 min., Add to 50 μL Z-DH5α