Bacterial Transformation Lab .pdf - Bacterial...

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Bacterial Transformation Lab QuestionsIntroduction Paragraph:In this lab, the effects of genetic modification will be observed usingE. Coli bacteria. A control group of unchanged DNA and DNA with ampicillin will also beobserved in the experiment. In the end we will be able to determine the changes geneticmodification played in the bacteria’s reproduction and phenotype. Observations from this lab canhelp analyze the results that other experiments involving GMOs and genetic engineeringproduce.Problem Statement:What is the effect of genetic modification on E. Coli using genes fromjellyfish and ampicillin related substances?Hypothesis: If the bacteria receives a ampicillin resistant trait then it will be able to reproduce ina high manner, because the ampicillin will not be able to kill it.Independent Variable: The plasmid in the DNADependent Variable: The amount of bacteria colonies that have grown.Control: The plate of LB that only used original bacteria and no genetic modification.Materials:-E. Coli bacteria-2 LB agar plates-2 LB/amp (ampicillin) agar plates-CaCl2-LB nutrient broth-Sterile inoculation loops-Pipettes-DNA plasmid-Microcentrifuge tubes for the DNAs and solutions-Ice bath beaker-Incubator-Aprons-Goggles-Ultraviolet light source-Timer-10% household bleachProcedure:1.Add 500 μl of CaCl2 into the Negative DNA centrifuge.
2.Using a toothpick, collect 15 colonies of E. coli bacteria and swirl it around in thecentrifuge made above.3.Then using a clean pipette, transfer 250 μl of the negative DNA to the positive DNAcentrifuge.4.Using the pipette again, transfer the positive DNA into the centrifuge containing 10 μlpGFP.5.Incubate these centrifuges in an ice bath for approximately 10 minutes.6.Remove tubes from ice bath and incubate at a temperature of 42°C for approximately 90seconds.7.After this, add 250 μl of “recovery broth” to both sets of centrifuges.

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Term
Fall
Professor
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Tags
Bacteria, DNA, Escherichia coli

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