221 Transformation of E. coli with pGAL™ (blue colony) - EDVOTEK

22EDVO-Kit # 221Transformation of E. coli with pGAL (blue colony)Pre-Lab PreparationsLabel (“Stripe”) the Plates6. Use a lab marker to "stripe" the sides of twenty (20) 60 x15 mmpetri dishes. This will provide an easy method of differentiatingbetween plates with ampicillin and plates without ampicillin.Instructor’s Guide• Open one sleeve of 20 plates and stack the plates neatly.• Start the marker at the bottom of the stack and move themarker vertically to the top plate to "stripe" the sides of the 20plates.• These plates will be used for medium with ampicillin.• Do not stripe the second sleeve of plates. These will be thecontrol plates.Pour the Plates (after the medium has cooled)7 . Thaw and add all of the X-Gal solution (Component E) to the moltenand cooled ReadyPour medium. Recap bottle and swirl to mix.Add reagents to mediumwhich has beencooled. Hot mediumwill cause reagentssuch as ampicillin torapidly decompose.8. Use a fresh 10 ml pipet and pipet pump to pour 10 unstriped plates,5 ml each. (See Quick Reference: Pouring Agar Plates.) Save thepipet for step 11.Quick Reference: Pouring Agar Plates• Use a sterile 10 ml pipet with a pipet pump to transfer thedesignated volume of medium to each petri plate. Pipet carefullyto avoid forming bubbles.• Rock the petri plate back and forth to obtain full coverage.• If the molten medium contains bubbles, they can be removed bypassing a flame across the surface of the medium.• Cover the petri plate and allow the medium to solidify.Duplication of this document, in conjunction with use of accompanying reagents, is permitted for classroom/laboratoryuse only. This document, or any part, may not be reproduced or distributed for any other purpose withoutthe written consent of EDVOTEK, Inc. Copyright © 1989, 90, 93, 94, 97, 98, 2000, 2005, EDVOTEK, Inc., all rightsreservedEVT 005077KThe Biotechnology Education Company ® • 1-800-EDVOTEK • www.edvotek.com