221 Transformation of E. coli with pGAL⢠(blue colony) - EDVOTEK
221 Transformation of E. coli with pGAL⢠(blue colony) - EDVOTEK
221 Transformation of E. coli with pGAL⢠(blue colony) - EDVOTEK
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<strong>Transformation</strong> <strong>of</strong> E. <strong>coli</strong> <strong>with</strong> pGAL (<strong>blue</strong> <strong>colony</strong>)EDVO-Kit # <strong>221</strong>Experiment Overview<strong>Transformation</strong> Experiment Flow Chart0.7 mlCells25 µlControl Buffer0.25 mlCells0.25 mlCells25 µlpGAL DNAThe ExperimentControlBufferIncubate on ice for10 minutesIncubate at 42°Cfor 90 secondsPlace on icefor 1 minuteAdd 0.75 ml recovery brothpGALDNAIncubate in 37°Cwater bath for 30 minutesFor optimal results, storecovered plates in the uprightposition after streakingto allow the cell suspensionto be absorbed by theagar. After approximately20 minutes, invert the platesfor overnight incubation at37°C.X-GAL/Control 1Incubateovernight in 37°Cincubation ovenAMP/X-GAL/Control 2Streak 0.25 ml <strong>of</strong> cellsuspension on platesIncubateovernight in 37°Cincubation ovenAMP/X-GAL/pGALX-GAL/Control 1AMP/X-GAL/Control 2AMP/X-GAL/pGALWhite Colonies No Colonies Blue ColoniesDuplication <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>outthe written consent <strong>of</strong> <strong>EDVOTEK</strong>, Inc. Copyright © 1989, 90, 93, 94, 97, 98, 2000, 2005, <strong>EDVOTEK</strong>, Inc., all rightsreservedEVT 005077KThe Biotechnology Education Company ® • 1-800-<strong>EDVOTEK</strong> • www.edvotek.com
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