To design: An experiment to test the hypothesis that B vitamin pyridoxine is an essential nutrient in mice. Introduction: Essential nutrients are the nutrients that cannot be synthesized by one’s own body but are required to carry out the normal physiological functions and must be provided through diet. There are six essential nutrients: carbohydrates, fats, protein, vitamins, minerals, and water.

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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?

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Question

Chapter 47, Problem 12TYU

Summary Introduction

To design: An experiment to test the hypothesis that B vitamin pyridoxine is an essential nutrient in mice.

Introduction: Essential nutrients are the nutrients that cannot be synthesized by one’s own body but are required to carry out the normal physiological functions and must be provided through diet. There are six essential nutrients: carbohydrates, fats, protein, vitamins, minerals, and water.

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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…

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…