IGEM:Virginia/2012/Notebook/Genetically engineered bacteriophage for diagnosis of whooping cough/2012/08/19
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Week of 8/19/12
• Digested T7 genome with SclI. No bands were present in the gel lanes containing the T7 genome. However, when the T7 genome that had been subjected to the same conditions as the digest, sans the addition of SclI enzyme, a smear appeared. Mostly of the smear was concentrated below the 100 bp marker, indicating that the genome was fragmented into very small pieces, either before or during the 4 hour long digestion at 50°C. This is odd as E. Coli (BL21) has been transformed before with the genome and produced intact phage. The genome, therefore, must have been intact for that experiment. Therefore, a gel extraction was run with the T7 genomne directly from the stock tube in the freezer using the QIAEX II kit. However, the kit failed as the solutions were vortexed too haed and too much QX1 buffer was added, resulting in failure.