User:Eric M. Walters/Notebook/Spring 2012/2012/03/13

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acsA-up-down, accBCDA digest Concentrations: acsA-up: 119 ng/μL acsA-down: 154 ng/μL accBCDA: 211 ng/μL Volumes to digest: acsA-up: 31 μL (all of it) acsA-down: 30 μL accBCDA: 19 μL Volumes of water (& 5 μL buffer 4): acsA-up: 13 μL acsA-down: 14 μL accBCDA: 24 μL Enzymes: acsA-up: 1 μL EcoRI-HF acsA-down: 1 μL BamHI-HF accBCDA: 1 μL EcoRI-HF, 1 μL BamHI-HF Run in the iGEM lab's 37 waterbath from 3:30-6:00, gel extracted. Ligation (50 μL)= Concentrations (up, acc, down): 48, 23, 63 ng/μL So reaction was: 3.5 up 21 accBCDA 3 down 5 buffer 5 ligase 16°C O/N