Haynes Lab:Notebook/Engineering PC-TFs/2015/04/13

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Summary Restriction Digest/Gel Verification, Concentration Reading Digest and Gel Verification Reagent Volume Expected: BL01 Ligation Colonies [XbaI/ApaLI] Forward insertion: 255, 288, 883, 1243, 2037, 3017 Reverse insertion: 288, 255, 1122, 1243, 2037, 2783 No insertion: 268, 883, 1243, 2783 BL09 Ligation Colonies [XbaI/ApaLI] Forward insertion: 255, 288, 883, 1243, 3977 Reverse insertion: 255, 288, 1243, 2082, 2783 No insertion: 268, 883, 1243, 2783 15 μL/lane; 1% agarose; Observed lengths: W1: Ladder W2: Bl09 TL V1 W3: Bl09 TL V2 W4: BL01 LSR V1 W5: BL01 LSR V2 W6: BL01 LSR V3 W7: BL01 LSR V4 Ladder DNA 3.0 μL 3.0 μL 2.0 μL 2.0 μL 1.0 μL 1.0 μL (From ladder 2 to ladder 6) 10X buffer 1.5.0 μL (for all) XbaI 1.0 μL (for all) ApaLI 1.0 μL (for all) dH2O 8.5 μL 8.5 μL 9.5 μL 9.5 μL 10.5 μL 10.5 μL Total 15 μL --> 37°C/ 15 min. Note: Punctured hole in Well 3. Wells 4 & 5 do not match with Wells 6 & 7. Rather, Wells 6 & 7 match with Wells 2 & 3. The tubes may have been mislabeled and Wells 6 & 7 could be Bl09 rather than Bl01. We will do another restriction digest and gel verification later this week to verify if that is the case.