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PCR Lab Report Revised by Tyler Nettelman

The purpose of the lab was to analyze DNA samples using PCR and gel electrophoresis to determine the frequency of Alu inserts in different genotypes. The student's initial results were inconclusive due to errors in measurement and calculations. Using a standardized data set provided by the teacher, the student determined the frequency of different genotypes in the class was: (-/-) = 21.1%, (+/+) = 29.3%, (+/-) = 49.7%. While the initial results were inconclusive, analyzing the standardized data helped the student better understand how PCR can be used to study DNA and ancestry.

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423 views

PCR Lab Report Revised by Tyler Nettelman

The purpose of the lab was to analyze DNA samples using PCR and gel electrophoresis to determine the frequency of Alu inserts in different genotypes. The student's initial results were inconclusive due to errors in measurement and calculations. Using a standardized data set provided by the teacher, the student determined the frequency of different genotypes in the class was: (-/-) = 21.1%, (+/+) = 29.3%, (+/-) = 49.7%. While the initial results were inconclusive, analyzing the standardized data helped the student better understand how PCR can be used to study DNA and ancestry.

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api-383435531
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© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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PCR Lab Report Revised Tyler

Nettelman
STEM 1
period
12/18/17

Purpose: The purpose of this lab was to track down the history of our DNA using
PCR and gel electrophoresis.

Hypothesis: We will determine the frequency of the Alu inserts by testing our
cheek cells using PCR. We will look at 3 different genome types: (+/+) (+/-) (-/-).
If we follow the steps to the procedure correctly then we should end up with
sequenced and replicated DNA segments.

Procedure: Before beginning the PCR lab we will be given a short but precise
understanding about agarose gel, Genotype, DNA, alleles, etc, if we do not
already have some experience with them in the previous years. We will also be
talking about the dos and the don'ts when in a lab atmosphere, because safety is
our utmost concern when we are in the lab. The following procedures apply. (In
accordance to BABEC ALU PCR 2017)

Data/ Observations:

First we inserted gel red into the lanes and allowed it to spread and stain our gel
red for 72 hours. We performed electrophoresis on our agarose gel (150V for 20
minutes). Lanes 1a and 2a were filled with 100 bp ladder. This allowed us to
compare the sequences in our DNA for sizing. All of the other lanes had 20 mL of
a 50 mL DNA/ 10 mL loading dye solution. I worked with lane 2c which worked
correctly. Of the other lanes, only 1g also worked correctly. All others did not
work correctly.
Gel after electrophoresis:

Lane Set Up:


1a 1b 1c 1d 1e 1f 1g 1h (1a=ladder)
2a 2b 2c 2d 2e 2f 2g 2h (1b=ladder)
Analysis:
Alu Repeat Frequencies

(+/+) 15 Students
(+/-) 10 Students
(-/-) 12 Students

Frequency (+) = # of (+) Frequency (-) = # of (-)


total # total #
= 40 = 34
74 74
= 0.541 or 54% = 0.459 or
46%

Genotype Frequencies: Equation:


p = (+) allele frequency P + 2pg + q = 1.0
2 2

q = (-) allele frequency (0.541) + 2(0.541)(0.459) + (0.459) =


2 2

1.0

Frequency of (-/-) = 0.211 = 21.1%


Frequency of (+/+) = 0.293 = 29.3%
Frequency of (+/-) = 0.497 =49.7%

Our groups data proved inconclusive. Because this was common throughout the
groups in the class, our teacher decided to make a fake set so we could analyze
every alu repeat within each set. We found different frequencies of various
alleles. These were the possible genes that were represented: (+/+) (+/-) (-/-).
(+/+) represents what an alu repeat looks like (alu inserts in both
chromosomes). An alu repeat is a short stretch of DNA originally characterized
by its action. (+/-) represents an alu insert in one chromosome. If you find out you
have (-/-) then sorry, you have no alu inserts.
After determining my results within the class set, I can conclude that my
hypothesis was correct. Because we were inexperienced, we did not follow the
steps correctly and therefore did not end up with sequenced and replicated DNA
segments. Our groups errors likely came from inaccurate pipet measurements
which caused the PCR reaction to not work. We had an error in our equation,
which led to an incorrect percentage of frequency. To improve this lab, it is
necessary to improve the accuracy of pipet measurements. This could be
accomplished by having the teacher check the measurements as we work, or by
comparing measurements with another group. To ensure that the equations are
correct, each person in the group should do them individually and compare
answers.
When we used the class set, I was able to retrieve a correct reading of my
DNA along with my alleles. However I would have been able to trace my own
ethnic roots and better my understanding about how DNA works if the original set
of data was done correctly. A further investigation of this lab could have been a
comparison of our class results to see who had identical alleles and were related
due to a common ancestor.

Conclusion:
The point of this lab was to find sequenced and replicated DNA segments. We
swabbed DNA from our cheek cells, performed electrophoresis on it, and
attempted to find Alu repeats in our DNA. When performed properly, this is an
efficient method for enzymatic amplification of specific DNA sequences, but due
to error, we were forced to use a fake set so we could analyze every alu repeat
within each set. Using that data we determined the following: the frequency of (-/-
) = 21.1%, (+/+) = 29.3%, and (+/-) =49.7% Incorrect measurements are a
common mistake, which can lead to inconclusive results. Working with the fake
set of data did allow us to determine that most of the fake class had an alu insert
in one chromosome. The PCR process can be used to aid students in a more
complete understanding of DNA and ancestral background.

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