Biology
Biology
12th Edition
ISBN: 9781260494570
Author: Raven, Peter
Publisher: MCGRAW-HILL HIGHER EDUCATION
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Chapter 8.7, Problem 1LO
Summary Introduction

To distinguish between: The way in which rubisco acts to make RuBP and explain the way in which it oxidizes RuBP.

Introduction: The citric acid is also called the Kreb’s cycle or tricarboxylic acid cycle. It is a series of chemical reactions used by all aerobic organisms to relate the stored energy by the oxidation of acetyl-CoA derived from fats, carbohydrates, and proteins into carbon dioxide and adenosine triphosphate.

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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…
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