Haynes Lab:Notebook/Engineering PC-TFs/2014/08/08

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Summary Concentration reading for gel purified PcTFs, ligation product minipreps. Gel verification of ligation attempt 3. Concentration Readings Gel Verification Reagent Volume Expected: BL01(Well: 2), BL05(Well:3-6) Forward insertion: 710, 7010. Reverse insertion: 3230, 4490. No insertion: 710, 4490. CMV/MV9 [XbaI/NruI] (Well: 7) 710, 4490 BL09 Ligation Colonies 1-6[XbaI/NruI] (Well: 8-13) Forward insertion: 710, 5921 Reverse insertion: 2141, 4490 No insertion: 710, 4490 15 μL/lane; 1% agarose; Observed lengths: W2(BL01Lig1): 5000, 700; W3(BL05Lig1): 5000, 700; W4(BL05Lig2): 5000, 700; W5(BL05Lig3): 5000, 700; W6(BL05Lig4): 5000, 700; W7(Empty CMV/MV9): 5000, 700; W8(BL09Lig1): 5000, 700; W9(BL09Lig2): 7000, 700; W10(BL09Lig3): 5000, 700; W11(BL09Lig4): 5000, 700; W12(BL09Lig5): 7000, 700; W13(BL09Lig6): 5000, 700; Ladder DNA(BL01x2, BL05x1, BL09x2, CMV/MV9x1) 2.0 μL 10X buffer 1.5 μL XbaI 1.0 μL NruI 1.0 μL dH2O 9.5 μL Total 15 μL --> 37°C/ 30 min. BL09Lig2&5 sent in for sequencing.