IGEM:British Columbia/2009/Notebook/Biosensor Sensitivity/2009/08/13

iGEM Project name 1

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assembled assemblies 2, 11, 12, 13, 27, 28, 22, 23, 24, 25 1. Digestion using standard UBC iGEM digestion protocol Prefix samples (cut with EcoRI and SpeI) BBa_J23100 Lock 1 (Amp) Key 1 (Amp) ASEM 21 Suffix samples (cut with XbaI and PstI) ASEM 17 ASEM 18 ASEM 19 ASEM 20 BBa_B0014 BBa_E0022 BBa_E0034 BBa_J31001 BBa_J61046 Backbone cut (cut with PstI and EcoRI) pSB1C3 pSB1A3 2. Ligation using standard UBC iGEM digestion protocol ligation into pSB1C3': i) inserts ASEM 02 (1.00μL) ASEM 11 (7.09μL) ASEM 12 (9.32μL) ASEM 13 (1.10μL) ASEM 27 (7.89μL) ASEM 28 (8.83μL) ii) reaction mixture vector: 2μL insert: please see volume in parathesis above water: 14μL - insert volume 10x buffer: 2μL ligase: 1μL -- ligation into pSB1A3': i) inserts ASEM 22 (10.42μL) ASEM 23 (9.51μL) ASEM 24 (9.94μL) ASEM 25 (10.41μL) ii) reaction mixture vector: 2μL insert: please see volume in parathesis above water: 14μL - insert volume 10x buffer: 2μL ligase: 1μL Overnight ligation in cold (ice) water bath transformed 10μL of each ligation mix

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