Agarose Gel Electrophoresis Overview

These flashcards cover key concepts related to agarose gel electrophoresis, including preparation, procedure, and principles.

What is the purpose of gel electrophoresis?

To separate DNA based on size and charge.

What three features are used to separate DNA in gel electrophoresis?

Size, charge, and shape.

What charge does DNA have?

DNA has a negative charge.

In which direction does DNA move in electrophoresis?

DNA moves toward the positive electrode (anode).

What is agarose gel made from?

Agarose gel is made from agarose powder dissolved in buffer.

Why do we use buffer instead of water in gel electrophoresis?

Buffer maintains a stable pH and provides ions for conductivity.

Which molecules move faster in the gel, large or small?

Small molecules move faster than large ones.

How is a 1.5% agarose gel prepared?

Combine 0.75 grams of agarose with 50 mL of TBE buffer.

What is the purpose of adding Gel Green dye?

To bind to DNA and fluoresce under UV light, allowing visualization.

What should be avoided when pouring the melted agarose solution?

Avoid air bubbles when pouring the agarose.

What does the 6X loading buffer contain?

Bromophenol Blue for color and Glycerol for weight.

What is the DNA ladder used for?

To estimate the size of DNA fragments in the gel.

At what voltage is the gel typically run?

The gel is typically run at 100V.

What should happen to the electrodes when power is applied?

Bubbles should form on the electrodes.

How long is the gel normally run during electrophoresis?

The gel is run for about 15-30 minutes.