221 Transformation of E. coli with pGALâ¢ (blue colony) - EDVOTEK

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18EDVO-Kit # 221Transformation of E. coli with pGAL (blue colony)Study QuestionsAnswer the following study questions in your laboratory notebook or ona separate worksheet.The Experiment1. Did you observe any satellite colonies? Why are the satellite, feedercolonies white?2. Why did the competent cells which did not receive DNA (control)fail to grow on the plates containing ampicillin?3. Why are there so many cells growing on the X-GAL plate? Whatcolor are they?4. What evidence do you have that transformation was successful?5. What are some reasons why transformation may be unsuccessful?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
Transformation of E. coli with pGAL (blue colony) EDVO-Kit # 221 19Instructor’s GuideNotes to the Instructor:Online Orderingnow availableVisit our web site forinformation aboutEDVOTEK's completeline of experiments forbiotechnology andbiology education.EDVO-TECH SERVICEMon - Fri1-800-EDVOTEK(1-800-338-6835)9 am - 6 pm ETImportant READ ME!Transformation experiments contain antibiotics which are used for theselection of transformed bacteria. Students who have allergies to antibioticssuch as penicilllin, ampicillin, kanamycin or tetracycine shouldnot participate in this experiment.Organizing and Implementing the ExperimentClass size, length of laboratory sessions, and availability of equipmentare factors which must be considered in the planning and the implementationof this experiment with your students.The guidelines that are presented in this manual are based on ten laboratorygroups consisting of two, or up to four students. The followingare implementation guidelines, which can be adapted to fit your specificset of circumstances. If you do not find the answers to your questionsin this section, a variety of resources are available at the EDVOTEK website. In addition, Technical Service is available from 9:00 am to 6:00 pm,Eastern time zone. Call 1-800-EDVOTEK for help from our knowledgeabletechnical staff.Technical ServiceDepartmentMon - Fri9:00 am to 6:00 pm ETFAX: (301) 340-0582web: www.edvotek.comemail: edvotek@aol.comPlease have the following information:• The experiment number and title• Kit Lot number on box or tube• The literature version number(in lower right corner)• Approximate purchase dateDay 1: (Prior to the Lab)• Prepare agar plates• Prepare LyphoCells (overnight incubation).• Dispense the DNA and control bufferDay 2: (Day of Lab Experiment)• Equilibrate water baths at 37°C and42°C; incubation oven at 37°C• Students transform cells and plate forovernight incubation.Day 3: (Day after Lab Experiment)• Students observe transformants andcontrols• Students calculate transformation efficiency• Follow clean up and disposal proceduresas outlined in the Laboratory Safety section.EDVOTEK - The Biotechnology Education Company ®1-800-EDVOTEK • www.edvotek.com24-hour FAX: (301) 340-0582 • email: edvotek@aol.comEVT 005077K
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221 Transformation of E. coli with pGALâ¢ (blue colony) - EDVOTEK