IGEM:Peking University/2008/Notebook/Group 1/2008/07/28

Group 1

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PKU_080728_G1_1 Picking up colonies-lacS/tet Enzyme cutting and gel running Miniprep Biobrick Report: Testing result-Xili's plasmid-pADH-lac1/2/S/tet Digestion-pADH-lac1/2-1/2-2/S/tet-ClaI-BamHI Gel running and testing Gel Picture: 080725-xili_plasmid_test-redigest lanes:1kb marker, 100bp marker, lac1, lac2-1, lac2-2, lacS, tet, lambda marker Transformation-xili-lac1-1/1-2/2-1/2-2/S-1/S-2/tet-1/tet-2(from two envelops) Picking up colonies|Miniprep|digestion(40min)|Gel running(140V) Gel Picture: 080728-xili_plasmid_test-redigest Lanes: lambda marker, {lac2-2-1, lac2-2-2, lac2-2-3, lac2-2-4, lac2-2-5, lac2-2-6, lacS-2-1}0727miniprep, {lac1-1-1, lac2-2-1, lac2-2-2, lac2-2-3, lacS-1-1, lacS-2-1}0726miniprep, 100bp marker Picking up colonies|Miniprep|digestion(90min)|Gel running(120V) Gel Picture: 080728-xili_plasmid_test-redigest-longer_time Lanes: {lac2-2-1, lac2-2-2, lac2-2-3, lac2-2-4, lac2-2-5, lac2-2-6, lacS-2-1}0727miniprep, lambda marker, {lac1-1-1, lac2-2-1, lac2-2-2, lac2-2-3, lacS-1-1, lacS-2-1}0726miniprep Conlusion: 1. The plasmids are right from the enzyme cutting result: only 700bp bands, no 1.4kb bands, showing the exact fragment we need. 2. The middle one looks nicer--nearly no smearing phenomena, which suggests the following aspects MAY be reasons: a, shorter enzyme cutting time (since BamHI has potential spark activity); b, higher gel running volt (time to run gel becomes shorter); c, new K buffer (other materials are totally the same, including the enzyme containers). 3. Longer time for digestion may generate some new bands, if not contamination in lane last but two of pic3...