Homework 6- transformation

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Purdue University *

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Biology

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Apr 3, 2024

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Homework 6- Due April 1 st Activity 1: Watch the following video and answer the questions below: https://www.labxchange.org/library/items/lb:LabXchange:255eb5d5:video:1 Please watch the above Transformation video , and answer the following questions: 1. What is the purpose of adding calcium chloride while preparing competent cells? ( 1 pt. ) 2. If heat shock is needed to open the cells so the plasmids can enter, why do we place the cells on ice for a few minutes after heat shock? (1 pt .) 3. Why do we label the LB plates on the bottom side? (1 pt .) 4. Why do you add 500 ul of SOC medium to the cells after transformation? (1 pt .) Activity 2: Please, read the Plating Bacteria and Selective Media article and answer the following questions: https://www.labxchange.org/library/items/lb:LabXchange:40c53f02:html:1 1.1 Imagine you transformed competent E.coli cells with pUC19 plasmid (in P+ tube) and set up a negative control transformation, where competent E.coli cells were mixed with water instead of plasmid (in P- tube). You plated the cells from both tubes on two types of media: LB and LB/Amp. Would you expect the formation of bacterial colonies on these plates for each transformation (Yes/No)? ( 4 pt .) Media/Plate Tube/Transformation P+ P- LB LB/Amp 2.2. If you transformed bacteria with a plasmid carrying the kanamycin (Kan) resistance gene and a gene for green fluorescent protein (GFP) under the control of an arabinose- dependent promoter, what media can you use to observe the bacteria grow? What media can you use to observe the bacteria glow? (Yes/No) (2 pts.) Transformed bacteria are able to: LB LB/Amp LB/Kan LB/Kan/ara Grow? Glow?
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