Human Anatomy & Physiology Plus Mastering A&P with Pearson eText -- Access Card Package (2nd Edition) (What's New in Anatomy & Physiology)
2nd Edition
ISBN: 9780134702339
Author: Erin C. Amerman
Publisher: PEARSON
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Chapter 19, Problem 2AYKA
Summary Introduction
Case summary:
The patient, Mr. Jackson, is facing severe bleeding for several days and is presented to the emergency room with a minor wound. He is suffering from hemophilia A, as examined by his medical history.
Characters in the case:
Mr. Jackson
Adequate information:
The patent is suffering from hemophilia A, which is a genetic disorder caused by a deficiency of factor VIII. This factor is involved in the intrinsic pathway of blood clotting.
To determine:
The effect of platelets given to Mr. Jackson to stop bleeding, as suggested by one of the co-workers.
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1)Which plate did you see purple/pink/blue bacterial cells? Why did you see this growth? Explain your answer in terms of transformation and plasmids?
2) Calculating Transformation Efficiency
For the +DNA/+Amp/+IPTG plate, record the following:
Number of transformants (colonies): _________________
Nanograms of plasmid DNA added: 50 ng
Final recovery volume: 0.50 mL
Volume plated: 0.25 mL
Transformation efficiency equation:
Transformation efficiency = Number of transformants / µg of DNA x Final volume at recovery (mL)/ volume plated (mL)
3) Using the equation above, calculate the transformation efficiency.
4) Describe the success of the transformation efficiency of this demo based on the calculation you did above?
1) Look at the ideal results. Were your predictions accurate, and how did they compare with your results?
2) You used aseptic technique during this lab. Why is it important to work in a sterile manner when working with bacteria in the lab?
3) Why are the cells incubated at 42°C?
Overview of Transformation Protocol
-Prepare competent bacteria for transformation:
Treat starter E. coli bacteria with CaCl2and Competent Cell Solution (CCS). Store on ice until transformation procedure.
Competent cells are cells that are likely to take up foreign DNA and be transformed. This step increases the likelihood that the E. coli cells will take up the introduced vector and be transformed.
-Transformation procedure:
Obtain two microcentrifuge tubes containing your competent cells. Label one tube +DNA and one -DNA.
Add CaCl2 to both tubes.
Add the transformation mix containing the plasmid DNA to the tube labeled +DNA. Do not add any plasmid DNA to the -DNA tube.
Incubate both tubes on ice for 10 minutes. Then, place both tubes in a 42\deg C water bath for 45 seconds. Replace the tubes in an ice bucket for 2 minutes.
Add recovery broth to both tubes.
Incubate both tubes in a 37 C water bath for 5 minutes.
Questions:
1) What differences would you expect to see between the…
Chapter 19 Solutions
Human Anatomy & Physiology Plus Mastering A&P with Pearson eText -- Access Card Package (2nd Edition) (What's New in Anatomy & Physiology)
Ch. 19.1 - Prob. 1QCCh. 19.1 - Prob. 2QCCh. 19.1 - 3. How does blood regulate temperature and...Ch. 19.1 - 4. What are the four main categories of plasma...Ch. 19.1 - Prob. 5QCCh. 19.2 - Describe the structure of a typical erythrocyte.Ch. 19.2 - Prob. 2QCCh. 19.2 - 3. Walk through the basic steps of...Ch. 19.2 - 4. What is the lifespan of an erythrocyte?
Ch. 19.2 - Prob. 5QC
Ch. 19.2 - Prob. 6QCCh. 19.2 - Prob. 7QCCh. 19.3 - How do granulocytes and agranulocytes differ?Ch. 19.3 - Prob. 2QCCh. 19.3 - Compare the two types of agranulocytes.Ch. 19.3 - 4. How do B and T lymphocytes differ?
Ch. 19.3 - 5. What does a monocyte become in the tissues?...Ch. 19.3 - Prob. 6QCCh. 19.3 - 7. How do the lymphoid and myeloid cell lines...Ch. 19.3 - 8. Where do T and B lymphocytes mature?
Ch. 19.4 - Prob. 1QCCh. 19.4 - Prob. 2QCCh. 19.5 - Prob. 1QCCh. 19.5 - Prob. 2QCCh. 19.5 - 3. How are platelets triggered to aggregate?
Ch. 19.5 - Prob. 4QCCh. 19.5 - How do the intrinsic/contact activation and...Ch. 19.5 - Prob. 6QCCh. 19.5 - Prob. 7QCCh. 19.5 - What is thrombolysis?Ch. 19.5 - Prob. 9QCCh. 19.5 - Prob. 10QCCh. 19.5 - Prob. 11QCCh. 19.5 - Prob. 12QCCh. 19.5 - Prob. 13QCCh. 19.5 - Prob. 14QCCh. 19.6 - Prob. 1QCCh. 19.6 - Prob. 2QCCh. 19.6 - Prob. 3QCCh. 19.6 - Prob. 4QCCh. 19.6 - Prob. 5QCCh. 19 - Prob. 1CYRCh. 19 - Prob. 2CYRCh. 19 - List the seven major functions of blood.Ch. 19 - Prob. 4CYRCh. 19 - Prob. 5CYRCh. 19 - Prob. 6CYRCh. 19 - Prob. 7CYRCh. 19 - Leukocytes are: a. nucleated cells that function...Ch. 19 - Prob. 9CYRCh. 19 - Fill in the blanks: Lymphocytes are derived from...Ch. 19 - Prob. 11CYRCh. 19 - Number the steps of hemostasis in order, putting 1...Ch. 19 - Prob. 13CYRCh. 19 - Prob. 14CYRCh. 19 - Prob. 15CYRCh. 19 - Prob. 16CYRCh. 19 - Prob. 17CYRCh. 19 - 18. Which antigens does a person with blood type...Ch. 19 - Prob. 19CYRCh. 19 - Prob. 20CYRCh. 19 - 1. Explain how blood, being a liquid, enables all...Ch. 19 - Prob. 2CYUCh. 19 - 3. The anticoagulant drug warfarin primarily...Ch. 19 - 4. Cirrhosis of the liver often reduces production...Ch. 19 - 1. A blood sample from your patient shows that she...Ch. 19 - Prob. 2AYKACh. 19 - Prob. 3AYKACh. 19 - Prob. 4AYKB
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- Overview of Transformation Protocol -Prepare competent bacteria for transformation: Treat starter E. coli bacteria with CaCl2and Competent Cell Solution (CCS). Store on ice until transformation procedure. Competent cells are cells that are likely to take up foreign DNA and be transformed. This step increases the likelihood that the E. coli cells will take up the introduced vector and be transformed. -Transformation procedure: Obtain two microcentrifuge tubes containing your competent cells. Label one tube +DNA and one -DNA. Add CaCl2 to both tubes. Add the transformation mix containing the plasmid DNA to the tube labeled +DNA. Do not add any plasmid DNA to the -DNA tube. Incubate both tubes on ice for 10 minutes. Then, place both tubes in a 42\deg C water bath for 45 seconds. Replace the tubes in an ice bucket for 2 minutes. Add recovery broth to both tubes. Incubate both tubes in a 37 C water bath for 5 minutes. Questions: 1)What is the selectable marker in this experiment? How…arrow_forwardBased on your results, which suspect's DNA best matches the DNA found at the crime scene?arrow_forwardIn oxidase test with Pseudomonas aeruginosa, the cell cultures on the slide turn colorless to be purple after tetra-methyl-p-phenylenediamine dihydrochloride (TMPD) is added. In the reaction, OTMPD is electron acceptor O cytochrome c is the electron source oxygen is terminal electron acceptor OH2 produced is electron donorarrow_forward
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