Transformation of E. coli with Green and Blue ... - Frederiksen
Transformation of E. coli with Green and Blue ... - Frederiksen
Transformation of E. coli with Green and Blue ... - Frederiksen
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26EDVO-Kit # 222<strong>Transformation</strong> <strong>of</strong> E. <strong>coli</strong> <strong>with</strong> <strong>Green</strong> <strong>and</strong><strong>Blue</strong> Fluorescent Protein PlasmidsPre-Lab PreparationsDay before the LabThis experiment requires preparation <strong>of</strong> isolated E.<strong>coli</strong> host transformationcolonies 16-24 hours before the laboratory experiment, so planaccordingly.Instructor’s GuideImportant: Do not prepare source plates more than 24 hours beforethe experiment. Old source plates will compromise the success <strong>of</strong> thetransformation experiment.E.<strong>coli</strong>Host cellsStreak cellsfor isolationonto sourceplates1. With a sterile loop, streak a small clump<strong>of</strong> E.<strong>coli</strong> cells from the slant <strong>and</strong> streak forisolation onto a source plate (see figures atleft).Do not use plates containing ampicillin.2. Repeat procedures as described for 4 additionalplates. You will have a total <strong>of</strong> 5source plates.LB Agar3. Incubate plates inverted overnight (16-24hours) in an incubation oven at 37°C.Incubate Source platesinverted overnight at 37°C.Duplication <strong>of</strong> this document, in conjunction <strong>with</strong> use <strong>of</strong> accompanying reagents, is permitted for classroom/laboratoryuse only. This document, or any part, may not be reproduced or distributed for any other purpose <strong>with</strong>out thewritten consent <strong>of</strong> EDVOTEK, Inc. Copyright © 1999, 2003, EDVOTEK, Inc., all rights reservedEVT 005077KThe Biotechnology Education Company ® • 1-800-EDVOTEK • www.edvotek.com
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