User:Wesley J. Houston/Notebook/MAP/2013/03/04

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3/4/13 Second ligation was successful. Assembly: (5xGal4 : [Spacer : HSVtk TATA]) : (K : AmCyan : AmCyan : NLS : STOP : [polyA2]) (KAH201)/(KAH182); AmCyan reporter gene for chromatin proteins. Assemblies Note, KAH201 is in vector V0120, so add 3200 bp to the size. KAH201/KAH182: (1) KAH201/(S/P)/203+3200 + (2) KAH182/(X/P)/1675 > Digests (Fermentas FD) Reagent Volume   30 μL/lane, 1% agarose; Ladder DNA (plasmid) 20 μL 10x buffer 3.0 enzyme 1 1.0 enzyme 2 1.0 dH2O 5.0 μL   30 μL --> 37°C/ ~15 min. > Measure conc.'s of gel purified fragments Sample OD260 260/280 ng/μL 1. KAH201 (S/P) 0.048 1.88 48.1 2. KAH182 (X/P) 0.008 1.93 8.1 > Ligations (corrected version, 3/04/13) Ligation Plate results (lig : neg crtl) 03/04/13 1. KAH201 (S/P)/3403, 25 ng + KAH182 (X/P)/1675, 26 ng 10 2. KAH201 (S/P)/3403, 25 ng 0