Lab Report 4 (Other (Not Listed) Sample)
Laboratory 4: Pipetting, DNA Extraction, PCR and Gel Electrophoresis
Learning Outcomes
• Learn pipetting practices, DNA extraction methods and PCR sample preparation.
• Develop familiarity with the equipment and materials used in biotechnology labs.
• Learn about the different components used in a PCR reaction and their function.
• Learn how PCR samples and prepared and how they would be loaded onto an agarose
gel.
• Analyze the results of a DNA agarose gel.
What you need to complete and the timeline:
1. Please note: There is no pre-lab quiz associated with Lab 4
2. Read the lab 4 required reading document.
3. Read Bioskill 8 and Section 20.2 in your textbook (p. 32-35; 414-415).
4. Read Bioskill 5 in your textbook (p. 26-29).
5. Read the definitions of the following terms: Deoxyribonucleic Acid, Nucleotide, Base
Pair, Polymerase Chain Reaction.
6. There are a series of videos associated with this lab. There will be questions associated
with the videos that you will answer via an online quiz.
You must watch the videos in the order in which they are presented on eClass. The
deadline to complete the questions for each video is shown below.
a. Lab 4 Introduction part 1 – No quiz questions associated with this video
b. Lab 4 introduction part 2 – No quiz questions associated with this video
c.
i. Using a micropipette
ii. Extraction of DNA from Cheek Cells
iii. Amplification of Extracted Cheek Cell DNA using PCR
iv. Running an Agarose DNA gel
Laboratory Report 4.
Student Name
University
Course Title
Professor Name
Date
Laboratory 4: Pipetting, DNA Extraction, PCR and Gel Electrophoresis
1 If a person is homozygous for the brown allele, how many bands will they see in the lanes for the blue and brown allele samples? (circle one) [1 mark]
Brown sample: 0 1 2 more than two.
Solution: The person will see 1 band.
Blue sample: 0 1 2 more than two.
Solution: The person will see 0
2 If a person is heterozygous, how many bands will they see in the lanes for the blue and brown allele samples? (circle one) [1 mark]
Brown sample: 0 1 2 more than two.
Solution: The person will see 1 band
Blue sample: 0 1 2 more than two.
Solution: The person will see 1 band
3 If a person is homozygous for the blue allele, how many bands will they see in the lanes for the blue and brown allele samples? (circle one) [1 mark]
Brown sample: 0 1 2 more than two.
Solution: The person will see 0 bands
Blue sample: 0 1 2 more than two.
Solution: The person will see 1 band
The image below is an example of the DNA agarose gel results. Each group (e.g. lanes labelled A) contains two samples, Bl=Blue and Br=Brown.
4 Interpret the results below as being homozygous blue, homozygous brown, or heterozygous. (2 marks).
lefttopA is Homozygous blue
B is Heterozygous
C is Homozygous brown
D is Homozygous blue
The sample lane listed as “LMW” denotes what is known as a Low Molecular Weight marker (see image below). This is a commercially available sample set of standard DNA fragments, measured in base pairs (bp). These standards of known size are used to determine the size of your PCR products (the DNA fragments you have created by PCR) by comparing the position of the sample bands with the bands of known size.
5 Based on the gels above and the image of the LMW marker below, extrapolate the size of the Blue and Brown eye colour alleles. (1 mark)
lefttopBlue eye color allele size (bp): 240 bp
Brown eye color allele size (bp): 190bp
Low Molecular Weight DNA Marker (New England Biolabs)
6 As you learned in labs 1-3, all experiments include controls. In this lab, a negative control was included along with the experimental
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