biogd281 lab outline

biogd281 lab outline - Chris Bostick BioGD281 Lab Outline...

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Chris Bostick BioGD281 5/13/2009 Lab Outline Purpose: To test lac- mutants isolated in experiment 2 to see if they are suppressed by one or more known amber suppressors. Materials: 1. 18 .2ml Pipettes 2. 1ml pipettes 3. 10ml pipettes 4. MacConkey’s Agar plates 5. Min + Glucose + Uracil + Tryptophan + Histidine + Streptomycin plates 6. Min + Glucose + Nalidixic acid plates 7. Min + Lactose + Uracil + Tryptophan + Histidine plates 8. Sterile Tooth Picks Strains Used in Experiment: Number Sex Genotype Properties Lac- HfrC Lac- Pro+, str(sen), nal(sen) CSH54 F- Deletion of lac-pro region, SupF Pro-, str(res) CSH55 F- Deletion of lac-pro region, SupE, nal(res) Pro-, nal(res) CSH56 F- Deletion of lac-pro region, SupD, nal(res) Pro-, nal(res) Res- resistant, sen- sensitive. Procedure: Step 1: Mix .2ml of the lac- mutant with 1.8ml of CSH54(mating F), 1.8ml CSH55(mating E), 1.8ml CSH56(mating D). Then incubate matings for an hour at 37 degrees Celsius. Step 2: Controls: (dilute to 10^-2 in distilled water) Plate .1 ml lac- on min + Strep and min + nalidixic acid Plate .1ml CSH54 on min + Strep Plate .1ml CSH55 on min + nalidixic acid
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