Electrophoresis and Micro-pipetting

Electrophoresis and Micro-pipetting

By Jasmine Jones

A biotechnology application that is used to produce a gel display of DNA or protein fragments compared for relatedness.​

What is Electrophoresis?​

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Multiple Choice

What is electrophoresis used for?

• To analyze the sugar in a gel

• To compare DNA and protein fragments

• To run electrical currents and break up DNA and proteins

• To view the structure of DNA and proteins

First a buffer solution is prepared and an agarose gel is casted. Then the gel is placed in a chamber with the buffer. The wells or holes is the gel are loades with samples. A power supply is connected to run a current through the buffer and the samples move to the positive end of the gel.

How is the application performed?​

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Multiple Choice

What needs to be added to the gel in the chamber for the electric charge to pass through?

• Sugar

• Distilled water

• A buffer solution

• Negatively charged DNA or protein

Multiple Choice

What charge does DNA have and to which charge will it move towards in the gel?

• Negative, Negative

• Positive, Positive

• Positive, Negative

• Negative, Positive

Multiple Choice

Explain the process of gel electrophoresis.

• Gel electrophoresis separates molecules based on size and charge.

• Gel electrophoresis separates molecules based on color and texture.

• Gel electrophoresis separates molecules based on taste and smell.

• Gel electrophoresis separates molecules based on temperature and density.

Multiple Choice

How does electrophoresis separate DNA fragments?

• By taste and smell

• By size and charge

• By temperature and pressure

• By color and texture

In order to load small amount of DNA into the well of the gel, a micropipette tool is used.

1. ​Read the plunger to find volume capacity

2. Adjust the volume to desired amount​

3. Click the plunger to the first point to draw fluids in

4. push the plunger completely down to expel fluids

5. ​eject the last tip using the tip eject button

Loading the Gel​

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Multiple Choice

What do you read to find the volume capacity of the micropipette?

• The volume reader

• The plunger

• The tip ejector

• The volume adjustment knob

Multiple Choice

When using a micropipettor, if you push to the second stop to fill it you will get ____________

• the correct amount of solution
• too much solution
• too little solution
• either too much or too little solution

Multiple Choice

The best tool to measure 146 uL (microliters) ______________.

• A PTP (plastic transfer pipette)
• A 5 - 50 uL micropipettor
• A 20 - 200 uL micropipettor

• A graduated cylinder

Multiple Choice

If the micropipette in this image is a P20, how many microliters could it measure?

• 0.98

• 9.8

• 98

• 980

Multiple Choice

If the micropipette in this image is a P200, how many microliters could it measure?

• 0.98

• 9.8

• 98

• 980

Multiple Choice

If the micropipette in this image is a P1000, how many microliters could it measure?

• 0.98

• 9.8

• 98

• 980

Multiple Choice

If the micropipette in this image is a P10, how many microliters could it measure?

• 0.98

• 9.8

• 98

• 980

Multiple Choice

This is the display from a P20 micropipette . How many microliters would be measured with this micropipette at this setting?

• 0.25

• 2.5

• 25

• 250

Multiple Choice

When dialing in a volume on a P-20 what should it read if you are putting in 9.2uL?

• 9

  2

  0

• 0

  9

  2

• 9

  0

  2

• 1

  9

  2

Multiple Choice

What tool would you use to measure 325 μl?

• P-20

• P-200

• P-1000

Multiple Choice

The best choice to measure 3 mL would be

• P-1000

• P-200

• 5 mL serological pipet

• transfer pipet

Multiple Choice

To what point do you push the plunger to draw up fluids into the tip?

• 1/4 of the way down

• All the way down

• 1/2 of the way down

• To the first point of resistance/first click

The metric system uses a base unit called the liter instead of gallons and ounces to measure volume. When you move down the metric ladder from the base unit, you are multiplying the amount by 10 or moving the decimal place to the right. When you move up the metric ladder from the base unit, you are dividing the amount by ten or moving the decimal to the left.​ Therefore, 1 liter is equal 1 million microliters, and 1 microliter are equal 0.000001 liters.

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In other words, ​numbers like microliters are even smaller when converted to bigger units like liters.

Converting metric units​

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Multiple Choice

What is smaller, a mililiter or microliter?

• mililiter

• microliter

Multiple Choice

How many microliters are there in liter of water?

• 1,000,000

• 1,000

• 100

• 100,000

Multiple Choice

How many liters of water is equal to 1 microliter?

• 0.0000001

• 0.0001

• 0.000001

• 1,000,000

1. Look at the migration distance of the mother's reference bands

2. Then look at the migration distance of the child's bands.

3. Since half of the DNA comes from the mother, the child's second band is related to the mother.

4. The correct father will have 50% of the DNA in common with the child as well​

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Answer who is the father on the next slide​

Interpreting Gel Electrophoresis​

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Multiple Choice

Who is the father of the child?

• alleged father #1

• Alleged father #2

• Alleged father #3