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Cut plasmids with BamH1 and HindIII 090711

2015-01-13

azim58 - Cut plasmids with BamH1 and HindIII 090711 When I cut PCANTAB5E with BamH1 and HindIII, the fragment should be a different size depending upon whether or not there was an scFv insert. Without an scFv insert: 2962-2236 = 726 With an scFv insert: 726+750 = 1476 BamH1 and HindIII both cut at 100% in Buffer 2 and both must be incubated at 1 hr. So I'll just perform the double digest at the same time in the same tube. I did not get the expected result (a cutout 700 bp fragment indicating that there is no insert). There could be several explanations. Maybe something went wrong with my double digest or maybe I have the wrong sequence for the plasmid and there is no HINDIII or BamHI site. It looks like at least one site was present since the plasmid was linearized. The single cut with BamHI linearized the plasmid so this site should be present. Sample Order: 100 bp GeneRuler Fermentas Single Cut Double Cut Uncut
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