A microbiome is an environment in which many species of microbes with their genes are assembled together to benefit the particular organism. To introduce the changes in the composition of a microbiome is the goal of microbiome engineering.

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Chapter 30, Problem 9TY

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Introduction: A microbiome is an environment in which many species of microbes with their genes are assembled together to benefit the particular organism. To introduce the changes in the composition of a microbiome is the goal of microbiome engineering.

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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…