6.3.3 Gel electrophoresis

• Gel electrophoresis separates molecules based on their size and charge
• What does gel electrophoresis analyze for DNA samples?

  Fragment sizes and mutations
• Smaller DNA fragments move faster through the gel matrix due to their smaller size
• What happens to smaller DNA fragments during electrophoresis?

  They migrate faster
• What forms the gel matrix in gel electrophoresis?

  Agarose powder
• The power supply in gel electrophoresis provides the electric field.


  True
• The running buffer in gel electrophoresis provides conductivity and maintains the pH for separation.


  True
• The power supply in gel electrophoresis provides the electric field
• What is gel electrophoresis used to separate?

  DNA, RNA, or proteins
• Steps in the gel electrophoresis procedure

  1️⃣ Gel casting

  2️⃣ Loading samples

  3️⃣ Applying electric field

  4️⃣ Staining/visualization
• DNA separation in gel electrophoresis uses an agarose gel and an electric field.

  True
• What creates the porous matrix in gel electrophoresis?

  Agarose gel
• Smaller DNA fragments migrate faster through the gel due to their smaller size
• What is the role of the running buffer in gel electrophoresis?

  Provides conductivity and pH
• The material used to form the gel matrix in gel electrophoresis is called agarose powder
• What is the purpose of a gel comb in gel electrophoresis?

  Creates wells for samples
• Loading dye is used to visualize DNA samples in gel wells during electrophoresis.


  True
• Match the gel type with its characteristic:

  Agarose ↔️ Large pore size

  Polyacrylamide ↔️ High resolution
• Gel electrophoresis is a technique used to separate molecules based on size and charge
• Smaller DNA fragments move faster through the gel matrix in gel electrophoresis.


  True
• Smaller DNA fragments move through the agarose gel matrix faster
• What is agarose powder used for in gel electrophoresis?

  Forms the gel matrix
• The electrophoresis chamber contains the gel and electrodes.


  True
• What is the purpose of loading dye in gel electrophoresis?

  Visualize samples in wells
• Step-by-step procedure for gel electrophoresis

  1️⃣ Prepare the gel

  2️⃣ Load samples

  3️⃣ Run the gel

  4️⃣ Stain and visualize

  5️⃣ Analyze results
• Without electrophoresis buffer, DNA separation is impossible in gel electrophoresis.


  True
• Steps to visualize DNA bands after gel electrophoresis

  1️⃣ Staining

  2️⃣ Washing

  3️⃣ UV transillumination

  4️⃣ Imaging
• Gel electrophoresis can be used to quantify RNA expression levels.


  True
• What is the purpose of a staining solution in gel electrophoresis?

  Makes DNA bands visible
• Steps for preparing and running a gel electrophoresis

  1️⃣ Prepare the Gel: Mix agarose powder with running buffer and heat. Pour into a casting tray with a comb to create wells.

  2️⃣ Load Samples: Add DNA samples mixed with loading dye into the gel wells.

  3️⃣ Run the Gel: Apply an electric current to the electrophoresis chamber.

  4️⃣ Stain and Visualize: Stain the gel with a dye to visualize DNA bands under a UV transilluminator.

  5️⃣ Analyze Results: Measure the distance traveled by the bands to estimate their sizes.
• What is visualized during the staining and visualization step of gel electrophoresis?

  Separated DNA bands
• What is the principle of DNA separation in gel electrophoresis based on?

  Size of DNA fragments
• Steps in the gel electrophoresis process

  1️⃣ Sample preparation

  2️⃣ Gel casting

  3️⃣ Electric field application

  4️⃣ Separation
• The running buffer in gel electrophoresis provides conductivity
• Match the equipment with its purpose:

  Power supply ↔️ Provides electric field

  UV transilluminator ↔️ Allows viewing stained gel

  Gel comb ↔️ Creates wells for samples
• What are the two primary roles of electrophoresis buffer?

  Conductivity and pH control
• The electrophoresis buffer maintains the appropriate pH
• Which staining solutions are commonly used to visualize DNA bands?

  Ethidium bromide and SYBR Green
• What is one application of gel electrophoresis in DNA analysis?

  Detect mutations