Experiment 3 Procedure - 5) Set up a master plate from...

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Chris Stanton March 26, 2009 Thurs Lab Rm 41 Experiment 3 Procedure 1) Mix 0.2 ml of the lac - with 1.8 ml of the CSH54 (F), CSH55 (E), and CSH56 (D). --Incubate each mating for one hour and 37°C 2) Set up controls and dilute to 10 -2 in distilled water. -plate 0.1 ml of lac - on min. + strep AND min. + nal -plate 0.1 ml of CSH54 on min + step -plate 0.1 ml of CSH55 and CSH56 on min + nal (separately) 3) Mix each mating on a vortex mixer for 60 seconds. 4) Plate 0.1 ml per plate from each mating mixture: -F: 10 -3 dilution on min + strep (glucose) -E: 10 -3 dilution on min + nal (glucose) -D: 10 -2 dilution on min + nal (glucose) -Incubate the plates (inverted) for 3-4 days at 37°C. Check each plate for colony growth after 24 hours and mark any small visible colonies when they appear.
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Unformatted text preview: 5) Set up a master plate from mating F on min + strep. Put 5-10 larger colonies on this master plate.-From the largest colonies, set up master plates from the matings of CSH55 and CSH56 on on min + nal plate. Put 5-10 colonies from mating E and D onto the plate.-Incubate all plates inverted at 37C. 6) Using the sterile toothpicks, patch each colony grown on the master plates to a single plate of min + lactose and a single plate of MacConkeys agar.-Include the control colonies on each plate and incubate plates at 37C. 7) Score the MacConkeys plates 12-20 hours after patching. Score the min + lactose plates 1-2 days after replication. 8) Make sure to record all results....
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This note was uploaded on 11/05/2009 for the course BIO 2810 taught by Professor Fox/nero during the Spring '07 term at Cornell University (Engineering School).

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