DNA Extraction and Bacterial Transformation Lab

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Questions and Answers

What is the primary purpose of adding the detergent/salt solution during DNA extraction?

To provide a nutrient source

To break down the cell membrane (correct)

To precipitate the DNA

To increase the temperature of the solution

Why is meat tenderizer added during the DNA extraction process?

To help dissolve lipids in the membrane

To lower the temperature of the solution

To remove cellular proteins attached to DNA (correct)

To precipitate DNA from the solution

What is the main function of ethanol in the DNA extraction process?

To make the cell membrane more permeable

To maintain the temperature of the solution

To precipitate the DNA and make it visible (correct)

To neutralize the negative charge of DNA

In the bioengineering lab, what is the initial step in preparing agar plates?

Mixing agar with boiling water Signup and view all the answers

What temperature is used to incubate the cells in the bioengineering lab?

37°C Signup and view all the answers

What happens during the 'heat shock' phase of transforming cells?

Cells are briefly exposed to high temperature to facilitate DNA uptake Signup and view all the answers

What is the purpose of the selective marker added to S plates?

To allow only transformed cells to grow Signup and view all the answers

What is the purpose of flipping the agar plates during incubation?

To prevent contamination from the incubator Signup and view all the answers

What is the primary goal of introducing foreign DNA into bacterial cells?

To observe genetic transformation and new traits expression Signup and view all the answers

What is the purpose of preparing a 2.0% agarose gel in electrophoresis?

To separate dyes based on size and charge Signup and view all the answers

What factor primarily influences how fast a dye migrates through the gel?

The size and charge of the dye Signup and view all the answers

Which of the following procedures is necessary to visualize the dye bands after electrophoresis?

Staining the gel, if required Signup and view all the answers

Which outcome indicates a successful separation of dye components in the gel?

A rainbow pattern appears on the gel Signup and view all the answers

What is the role of the electric current in the electrophoresis process?

To drive the migration of dyes based on charge and size Signup and view all the answers

How does Crystal Violet behave under certain conditions during electrophoresis?

It travels towards the negative electrode Signup and view all the answers

Which aspect of the transformed cells is being observed after incubation?

Presence of new traits like antibiotic resistance Signup and view all the answers

What is a common application for the technique of electrophoresis in research and diagnostics?

Quality control in pharmaceuticals Signup and view all the answers

What happens to larger or less negatively charged dyes during electrophoresis?

They move slower than smaller or more negatively charged dyes Signup and view all the answers

Study Notes

DNA Extraction Lab

Strawberry DNA Extraction: Strawberries are ground to release cell contents.
Detergent/Salt Solution: Breaks down cell membranes (lipids), neutralizes DNA's negative charge, allowing strand formation.
Meat Tenderizer (Enzyme): Breaks down proteins attached to DNA.
Ethanol: Precipitates DNA, making it visible. Cold ethanol prevents DNA from dissolving.

Bioengineering Lab: Bacterial Transformation

Agar Plates: Prepared with LB agar, adding a selective marker (pill) to certain plates.
Growing Blank Cells: Bacterial cells are streaked onto agar plates (NS, S-) to culture.
Making Cells Competent: Cells are prepared using cold temperature for easier DNA uptake.
Transforming Cells: Foreign DNA is added to competent cells; a heat shock step facilitates DNA entry.
Recovering Cells: Cells are placed in LB media to recover and express the introduced DNA.
Plating Cells: Transformed cells are spread on selective plates.
Incubating Cells: Cells are grown at 37°C, allowing comparison between transformed and non-transformed cells.

Electrophoresis Lab: Dye Separation

Gel Preparation: Agarose gel is made for separating dyes based on size and charge.
Dye Loading: Dye samples are loaded into designated wells. Precise volumes (10 µL) are used.
Electrophoresis: Electric current separates dyes in the gel based on size and charge. Smaller or negatively charged dyes migrate faster.
Gel Visualization: Stained or unstained gel viewed under UV for separation analysis. The process creates a distinguishable pattern.
Real-world Application: Commonly used for DNA, RNA, and protein analysis in research, quality control, and forensics. An electric current is employed to separate the dyes based on various properties. Crystal violet's movement to the negative side might indicate a positive charge.

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Description

This quiz covers the methods and processes involved in DNA extraction from strawberries and bacterial transformation using competent cells. You will learn about the chemicals and steps necessary to isolate and manipulate DNA. Test your knowledge on laboratory techniques crucial for bioengineering.