Making a series of dilutions, you transfer 40 ul (microliters) from a Bacteriophage-T4 stock to 4 ml (4000 ul) of Tryptone Soy Broth (TSB) in tube A.From tube A, you mix and transfer 40 ul to 4 ml TSB in tube B. You repeat this process from Tube B to Tube C, and then from Tube C to tube D. You mix100 ul (0.1 ml) of the Tube D phage dilution with 100 ul of a dense culture of susceptible E. coli in molten soft agar, and plate to grow overnight at 37*C.The next morning, you count 237 plaques from your plating of the tube D phage dilution. What was the original concentration of phage particles inyour Bacteriophage-T4 stock culture?O 237 x 10^10 pfu/ml2.37 x 10^8 pfu/ml2.37 x 10^11 pfu/ml2.37 x 10^5 pfu/mlO 23.7 x 10^8 pfu/ml

Bacteriophages are viruses that infect bacteria. When the bacteriophages are added to the bacterial…

Answered: Making a series of dilutions, you…

Biochemistry

9th Edition

ISBN:9781319114671

Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.

Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.

Chapter1: Biochemistry: An Evolving Science

Section: Chapter Questions

Problem 1P

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A plaque assay is performed beginning with 1 mL of a solution containing bacteriophages. This solution is serially diluted 3 times by combining 0.1 mL of each sequential dilution with 9.9 mL of liquid medium. Then 0.1 mL of the final dilution is plated in the plaque assay and yields 12 plaques.What is the initial density of bacteriophages in the original 1 mL? Enter your answer to two significant figures ( for example: 1.1 * 10^2)
Imagine that you are a student in Alfred Hershey and Martha Chase’s lab in the late 1940s. You are given five test tubes containing E. coli bacteria infected with T2 bacteriophages that have been labeled with either 32P or 35S. Unfortunately, you forget to mark the tubes and are now uncertain about which were labeled with 32P and which with 35S. You place the contents of each tube in a blender and turn it on for a few seconds to shear off the phage protein coats. You then centrifuge the contents to separate the protein coats and the cells. You check for the presence of radioactivity and obtain the following results. Which tubes contained E. coli infected with 32P-labeled phage? Explain your answer. Tube number Radioactivity present in 1 Cells 2 Protein coats 3 Protein coats 4 Cells 5 Cells
A plaque assay is performed beginning with 1 mL of a solution containing bacteriophages. This solution is serially diluted three times by combining 0.1 mL of each sequential dilution with 9.9 mL of liquid medium. Then 0.1 mL of the final dilution is plated in the plaque assay and yields 17 plaques. What is the initial density of bacteriophages in the original 1 mL?
Imagine that you are a student in Alfred Hershey and Martha Chase’s lab in the late 1940s. You are given five test tubes containing E. coli bacteria infected with T2 bacteriophages that have been labeled with either 32P or 35S. Unfortunately, you forget to mark the tubes and are now uncertain about which were labeled with 32P and which with 35S. You place the contents of each tube in a blender and turn it on for a few seconds to shear off the phage protein coats. You then centrifuge the contents to separate the protein coats and the cells. You check for thepresence of radioactivity and obtain the following results. Which tubes contained E. coli infected with 32P-labeled phage? Explain your answer.
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Dr. Wakefield would like to isolate recombinant plasmids from her bacterial culture using the alkaline lysis method. She is planning to use the chemicals as listed below: Solution I: 50 mM glucose, 25 mM Tris-Cl (pH 8.0), 10 mM EDTA (pH 8.0) Solution II: ??? Solution III: 5 M potassium acetate, glacial acetic acid, de-ion water Ethanol 70% (v/v) Isopropanol TE-RNAase pH 8.0 (i) (ii) (iii) Based on the chemical list above, state the content(s) of Solution II. Explain the functions of Solution II described in Q3 a) (i) in plasmid isolation. What is the role of alcohol precipitation conducted after the plasmids are obtained at the end of the procedure? Discuss the roles of ethanol and salt in alcohol precipitation step.
We need to prepare a stock solution of medium for your culture cells, which usually includes liquid salt solution and bovine serum. Our liquid salt solution is supplied in a 50X concentration, and we need to dilute it to 1X for use. We also need to add 75% fetal bovine serum for a final concentration of 15%. How would we make up 0.80 liters of this culture media using water as our solvent?
You take 10 ml of a stock solution, which is at a concentration of 1000 phage/ml, and dilute it to a total of 100 ml. From the resulting solution you take 5 ml and dilute it to 25 ml, and from the latter you take 5 ml and make a total of 20 ml. a) It will be possible to know how many bacteriophage particles there will be in 1 ml of the last solution b) What is the dilution factor in each step, in the same order in which the dilutions are made? c) What is the total serial dilution factor?
You perform a plaque assay with your bacteria and serial dilutions of your bacteriophage, , and then count the plaques. In the 10-5 dilution you have 318 plaques. In the 10-6 dilution, you count 32 plaques, and in the 10-7 dilution, there are 3 plaques. What is the approximate PFU/cell of your sample? 3.2 x 105 318 3.2 x 107 O 3.2 x 106
You overexpressed (asked bacteria to make a lot of) an 80 kDa protein carrying a 6X-His tag and purified it on a nickel column. You collect frac ons from your purifica on and analyze them using SDSPAGE. The results are shown in image attached. a) Interpret the results from the gel. What is wrong? (see image attached) b) Identify 2 possible reasons why this problem is occurring, and for each reason, suggest an experimental modifica on you can do to troubleshoot.
The following image is of an agarose gel. If DNA samples were loaded to this gel and the electrophoresis experiment was started, explain what would happen and why.
A high cell density culture of recombinant E. coli was carried out according to the following strategy:-Step 1: single batch with exponential growth until 98% conversion of the substrate, starting from V0= 4.0 L, S0=50 g/L/ X0= 1.0 g/LStep 2: batch fed with exponential flow (SF-800 g/L, μ= 0.1 h-1) until reaching X= 50.9 g/L;Step 3: batch fed with constant flow (F= 0.1 L/h) for 4 hours (induction phase with IPTG)Note: consider that the quasi-steady state is reached in both fed-batch stages.Extra data: YX/S = 0.4 gx/gs; μmax= 0.25 h-1; Ks== 1.0 g/L a) What was the cell concentration reached at the end of step 1?b) For step 3, considering that the substrate concentration in the feed was 1/4 of that used in step 2, what was the concentration of cells reached at the end of step 3?C) In terms of cell productivity, which of the three phases of cultivation was the most productive?

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