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

Transformation of E. coli with pGAL (blue colony) EDVO-Kit # 221 13Transformation of E.coliPlating the cellsPlating cells from the tube labeled "Control":12. Use a fresh, sterile 1 ml pipet to transfer recovered cells from thetube "Control Buffer" to the middle of the following plates:• 0.25 ml to the plate labeled X-GAL/Control 1• 0.25 ml to the plate labeled AMP/XGAL/Control 213. Spread the cells over the entireplate with a sterile inoculatingloop.14. Cover both control plates and allowthe liquid to be absorbed.The ExperimentSpread cells inone directionSame plate -spread cells 90°to first directionTo avoid contamination when plating, do notset the lid down on the lab bench -- Lift thelid of the plate only enough to allow spreading.Be careful to avoid gouging the loopinto the agar.Plating cells from the tube labeled "pGAL DNA"15. Use a fresh, sterile 1 ml pipet to transfer recovered cells from thetube "pGAL DNA" to the middle of the following plate:• 0.25 ml to the plate labeled AMP/X-GAL/pGAL16. Spread the cells with a sterile inoculating loop.Reminder:Follow properproceduresfor disposal ofcontaminatedmaterials.17. Cover the plate and allow the liquid to be absorbed (approximately15-20 minutes).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