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Unformatted text preview: Nicholas Sommariva Biology 141 Lab Glen Meades Materials and Methods for Transformation and Isolation of -Lactamase E. Coli cells were divided in half (100 L in each micro tube) and then 2 L of pBR322 were added to one tube and 2 L of TE buffer (control to ensure same environment in each sample) to the other using a pipette. Tubes were placed on ice for 20 minutes and then sat at room temperature for 10 minutes. One mL of Luria broth was added to each tube and placed on a shaker at 37C for 20 minutes. Bacteria containing the plasmid (transformed bacteria) and the buffer were then added to plates with a Luria agar. Cells were transferred to the plates in 2 L, 20 L and 200 L increments. Twelve plates were separated into having normal agar (control group) or having antibiotics in the agar (ampicillin) and both sets of bacteria were applied to the plates accordingly, that is all six positive plates (with ampicillin) and all six negative plates (with out antibiotics) received...
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This note was uploaded on 10/17/2011 for the course BIO 141 taught by Professor Dr.cafferty during the Fall '11 term at Emory.
- Fall '11