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Exgerim^ent Procedures

EDVO-Kit # 222 Transformation of E. coli with pFluoroGreen™ and pFluoroBlue^rM

Transformation of E.coli

PREPARING PLATES FOR INCUBATION

20.    Stack your group's set of plctes on top of one another and tape them together. Put your initials or group number on the faped set of plates.

The plates should be left in the upright position to allow che celi suspension to be absorbed by the agar.

21.    Place the set of plates in a safe place designated by your instructor.

22.    After Ihe cel! suspension is completeiy absorbed by the agcr, you or your insrructor will place the plates in the inyerted position (agcr side on top) in a 37°C bacteria! incubation oven for overnight incubation (15-20 hours).

The plates are inverted to prevent condensation on the lid, which could drip onto the culture and interfere with experimental results.

V!EWING PLATES AFTER INCUBATION

23.    Darken the room and use a long wave U.V. light to vi$uaiize the transformed cells that will glow as green or blue due to the exprcssion of the green or blue fluorescent proteins.

To visualize che fluorescent colonies, the long wave U.V. light (EDVOTEK cat. tt 969 recommended) can be held underneath the plates in a darkened room.

24.    Proceed to detcrmining the transformation efficiency.

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^    \    C V Dupllcacion of this document. in conjonction with use of accompanying reagents. is peiTnitced for classroom/

C & V W i C si\® laboratory use only. This document. or any part. may not be reproduced or discributed for any other purpose ^r _/    / wichout che writcen consent of EDVOTEK. Inc. Copyright © 1999.2003 EDYOTEK. Inc.. al! rights reserved.

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