Theoretical Data Following the modified protocol for the isolation of Escherichia coli bacteriophage of Encabo (2018) presented below, compute for the pfu/ml of the chloroform-treated lysate. 1ml 1ml 1ml 1ml 1ml Chloroform-treated lysate Dilution 10-³ [[ ]]] 9ml 104 10-4 10-5 9ml 10-1 9ml 10-² Empty sterile tubes Incubate inside ref for 15-20min + 0.1 ml lysate-E.coli mix 9ml 103 ↓↓ 0.1 ml DO 0.1 ml 9ml 10.5 0.1 ml +0.5 ml E. coli AA Incubate O/N at 35°C, observe for plaques Molten soft agar overlay ↓ OO B. Quantification of the Infective E. coli other Bacteriophage Particles in the Raw Sewage Sample Volume of Stock Plated (in ml) Bottom agar No. of Plaque-Forming Units (pfu) Ave. A B 254 265 132 110 11 23

Theoretical Data Following the modified protocol for the isolation of Escherichia coli bacteriophage of Encabo (2018) presented below, compute for the pfu/ml of the chloroform-treated lysate. 1ml 1ml 1ml 1ml 1ml Chloroform-treated lysate Dilution 10-³ [[ ]]] 9ml 104 10-4 10-5 9ml 10-1 9ml 10-² Empty sterile tubes Incubate inside ref for 15-20min + 0.1 ml lysate-E.coli mix 9ml 103 ↓↓ 0.1 ml DO 0.1 ml 9ml 10.5 0.1 ml +0.5 ml E. coli AA Incubate O/N at 35°C, observe for plaques Molten soft agar overlay ↓ OO B. Quantification of the Infective E. coli other Bacteriophage Particles in the Raw Sewage Sample Volume of Stock Plated (in ml) Bottom agar No. of Plaque-Forming Units (pfu) Ave. A B 254 265 132 110 11 23

Biology: The Unity and Diversity of Life (MindTap Course List)

14th Edition

ISBN:9781305073951

Author:Cecie Starr, Ralph Taggart, Christine Evers, Lisa Starr

Publisher:Cecie Starr, Ralph Taggart, Christine Evers, Lisa Starr

Chapter8: Dna Structure And Function

Section: Chapter Questions

Problem 4DAA: HersheyChase Experiments The graph shown in FIGURE 8.5 is reproduced from an original 1952...

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Calculate the amount of GFP in grams produced in the 25-L fermentation used to generate cells for your homogenization lab. To do this problem, recall that for the homogenization step, E. coli cell paste was suspended in Tris buffer, the resulting cell suspension was homogenized. For this calculation, assume that the concentration of GFP in the clarified lysate is 2.0 mg GFP/mL solution. The concentration of cells in the 25-L fermentation was 8% by volume.
In a nutrient medium that lacks histidine, a thin layer of agar containing ~109 Salmonella typhimurium histidine auxotrophs (mutant cells that require histidine to survive) produces ~13 colonies over a two-day incubation period at 37 °C. How do these colonies arise in the absence of histidine? The experiment is repeated in the presence of 0.4 μg of 2-aminoanthracene. The number of colonies produced over two days exceeds 10,000.What does this indicate about 2-aminoanthracene? What can you surmise about its carcinogenicity?
Q. You have now carried out the Somogyi Nelson determination of reducing sugars and measured the absorption spectra of Tube 9. What is the optimum wavelength you should use to monitor the Somogyi Nelson Assay why? Table 1._SOMOGYI-NELSON DTERMENATION OF REDUCING SUGARS Tube 1 2 3 4 5 6 7 8 9 10 D-Glucose (160 µg/mL), mL 0 0 0.125 0.125 0.25 0.25 0.375 0.375 0.5 0.5 H2O, mL 0.5 0.5 0.375 0.375 0.25 0.25 0.125 0.125 0 0 D-Galactose (160 µg/mL), mL - - - - - - - - - - D-Arabinose (160 µg/mL), mL - - - - - - - - - - Glucose-Galactose Mixture, mL - - - - - - - - - - Absorbance (710 nm) 0.049 0.051 0.138 0.151 0.271 0.26 0.382 0.395 0.454 0.444 Absorbance - blank 0 0 0.088 0.101 0.221 0.21 0.332 0.345 0.404 0.394 Final [Reducing sugar]…
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The purification of cytochrome C begins with 1) yeast homogenization using a bead beater in the presence of BME (a reducing agent) and a protease inhibitor from approximately 900 grams of Baker’s yeast. Then, 2) insoluble cell contents were removed by centrifugation at 4,000 x g for approximately ten minutes. The ruptured cells (lysate) after centrifugation had a total volume of 0 mL and a 1.0 mL aliquot was set aside for further analysis. The following data was obtained from the 1.0 mL aliquot to quantify the protein amount and purity: The absorbance at 410 nm of the aliquot was 0.460 (1 cm pathlength). The absorbance at 595 nm from a 1.0 mL Bradford Assay solution that was diluted by 250-fold from the aliquot was 0.681 (1 cm pathlength). Using the information given, Calculate the total protein amount in mg from the absorbance at 595 nm. Calculate the cytochrome C amount in mg from the absorbance at 410 nm using Beer’s Law.
Antibiotics that target bacterial molecules not previously exploited are desperately needed. One such target is the protein FtsZ. The small molecule 3-methoxybenzamide (3-MBA) is known to inhibit FtsZ in Bacillus subtilis but is not bacteriocidal. Nonetheless, researchers reasoned that 3-MBA offered a good starting point for the synthesis of a molecule that might be a potential drug candidate. Over 500 3-MBA analogues were synthesized and screened; one called PA190723 was extremely potent in its capacity to bind FtsZ and inhibit bacterial growth. In fact, when used in a mouse model, PA190723 was bacteriocidal against methicillinand multidrug-resistant Staphylococcus aureus. What makes FtsZ a good drug target? What preliminary information about 3-MBA would be helpful if you were designing the 3-MBA analogues? As these researchers move forward with clinical (human) testing, what other parameters and outcomes must be assessed besides the bacteriocidal activity of PA190723?
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