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Digest

Plasmid was digested with StuI according to NEB protocol for two hours instead of one

Gel

From Left to right: Hyperladder, hEF1a:CD79A-pTet:RFP-Gal4VP16 sample 1, 2, 3, 4, 5

Expected overlaid bands at 4kb and at slightly over 1.5kb

Image Added

used NEB two log ladder. Bands appear to be at 4kb and around 1.5kb on all samples. Smearing on sample 3 appears to show contamination in sample three as predicted in the miniprep.