Matthew Sargent

PD-1,2
9/26/17
Lab Write Up

Purpose: The Purpose of this lab was to give us more lab experience and to hopefully track the

history of our own DNA and learn where we came from. We tried to do this by using PCR and

electrophoresis to test and analyze the data.

Hypothesis: If I perform PCR and gel electrophoresis then it will show my ancestry comes from

English or Scottish descent.

Procedure: In accordance to BABEC ALU PCR 2017.

Data/Observation: In this experiment there were not many things that I could observe or take

data from because our data was inconclusive. I did notice that during the experiment the DNA

would sink to the bottom of the test tubes after centrifuging because of the DNAs higher density

than the liquid.

In this lab we used a 2% agarose gel that ran at 150 v for 20 minutes and stained using gelred

for 72 hours. Lane A1 and B1 have a 100 bp ladder lane A2-8 and B2-8 have a 20ml

(microliters) of a 50ml DNA/ 10ml loading dye solution. My column

Analysis and Discussion:

Genotype Number of Students Number of + Alleles Number of - Alleles

++ 15 30 0

+- 10 10 10

-- 12 0 24

Total 37 40 34

Allele Frequencies:

40 total + alleles

40/74=.541

.541= + frequency

34 total - alleles

34/74=.459

.459=- frequency

.541^2=.293

.293(37)=11 people +/+

.459^2=.211

.211(37)= 8 people -/-

2(.541)(.459)=.497

.497(37)=18 people +/-

Genotype Expected Genotype Total Number of Expected Number of

Frequency Students in Class Students with
Genotype
 +/+ .293 15 11

+/- .497 10 18

-/- .211 12 8
This data can neither support or go against my hypothesis because this data does not pertain to

our class. In our class we did not receive data so we could not make accurate calculations that

would support or deny our hypothesis. Some errors in our experiment could have been that we

did not pipet the correct amount of DNA or other fluids that were added to our saliva. Another

error that could have occurred was the heat or cold that was applied to our solutions for a

lengthy period of time. The temperature might have been off or the time in the varying

temperatures might have been wrong which could also cause error in the experiment. Some

ways we could improve the lab is by getting more familiar and more experienced with pipetting

before doing it for the real thing. This could improve our accuracy and results with the DNA

significantly due to the higher quality pipet work. Another thing we could improve about this

experiment is the quality and sterilization of tools before use in the lab. This might be hard to

accomplish due to a limited budget but good quality and sterile tools could greatly increase the

chance for success.

Conclusion: Working with pipettes to get accurate and clear data is very difficult. This

experiment involved taking DNA from our mouth cells and using PCR to replicate it. We then

took the DNA and mixed it with a series of other liquids and temperatures finally putting it on

agarose gel electrophoresis, with a goal of trying to find Alu repeats in our DNA and compare

them to other peoples alu repeats. Trying to find the place where our ancestors came from.

Using pipettes, if used correctly, can make many labs more precise with accurate

measurements of liquid. However, in this case where my class and I had only had a class period

to practice using them they became our obstacle. For example, when combining our DNA from

our cheek cells into the other liquids,we had to first centrifuge the DNA and then accurately

transport it into another test tube. This became an issue after inaccurate measurements lead to
faulty end results. Another example of a pipette dilemma is when we moved our thought to be

alu repeats onto the agarose gel. By this time I had figured out how to accurately pick up the

right amount of liquid in the pipette but this time we had to put this liquid into skinny wells that

would show results of our repeats. With minimal amounts of practice my classmates and I

struggled with putting the repeats into the wells causing them to spill out. This also could have

been one of our main problems that made our lab get no results. Pipettes are fabulous tools in

labs with advanced and experienced scientists but without proper practice they could become a

hassle that could potentially ruin the results of your experiment.