Bio lab 2 report - the tube labeled “-” 5 Incubate on...

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Title Simi Simon, March 22, 2007, Stephen Huff Abstract The purpose of this experiment is to select which cells are transformants and which cells are not by placing them on agar plates containing ampicillin. The cells of the bacteria called E. coli are formulated to take up the plasmid pAMP that contains a gene for ampicillin resistance. Then the cells are allowed to grow on the agar plates. The cells that survive and grow have been transformed by the plasmid and the cells that die have not been transformed. Materials and Methods 1. The first measure of our experiment is to thaw the competent cells on ice 2. Label one clear eppendorf tube with a “+” and the other with “-” sign. Write your bench letter on each tube. 3. Place 50 µl of competent cells into each microtubule. 4. Add 10 µl of pAMP plasmid into the tube labeled “+” and 10 µl of TE buffer into
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Unformatted text preview: the tube labeled “-”. 5. Incubate on ice for 15 minutes. 6. Label two LB Agar plates one as “LB+” and the other as “LB-”. Label two LB/Amp Agar plates one as “LB/Amp +” and the other as “LB Amp(-)”. Write your bench letter on each plate. 7. Heat-shock the cells at 42 C for 90 seconds. 8. Immediately return cells to ice for 2 minutes. 9. Add 200µl of Luria broth to each rube. Mix gently by tapping it with your finger. 10. Incubate at 37 c for 45 minutes. 11. Place 70 µl of “+” cells on center of “LB(+)” Agar plate and spread using a sterile L shaped glass rod. 12. Repeat step 11 on “LB/Amp(-)” 13. Allow plates to set for 5 minutes. 14. Tape plates for the thoe whole class together and label with TA’s name. 15. Incubate inverted plates overnight at 37 C....
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This note was uploaded on 04/14/2008 for the course BIOL 1162 taught by Professor Anamedrano during the Spring '08 term at University of Houston.

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