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Purpose:
The purpose of the lab was to gain lab experience and to learn about PCR. We
learned about PCR through using our DNA to find our alu repeats.
Hypothesis:
If I follow this procedure correctly then I will be able to see my alu repeats, which
Procedure
We followed BABECs procedures on finding your own alu repeats using PCR.
To be safe in the lab, we tried to stay quiet so we could hear directions. We also made
sure we changed our pipet tip or each person so we would not have our DNA in
Data/ Observations:
We made a 2% agarose which we ran at 150 volt for 20 minutes. We stained it
using gel red and we ran it for 72 hours. In lanes 1A and 2A we had our ladder which
appears orange-pink in the picture. Each gel slot (1B-H and 2B-H) had a 20mL sample
of our 50mL DNA with 10 uL loading dye added to the solution. My sample was in lane
On the first day of the lab, we had to swish a salt rinse inside of our mouth to
extract cheek cells which hold DNA. The salt water tasted very bad and I dont think I
was rinsing it long enough so that could be why my lad did not go so well. I also notice
how small the Chelex beads were and how careful I had to be to make sure I did not
On the second day I saw the importance of keeping the PCR tube on ice
because it had to stay at 4C in order for the lab to be accurate. The tiny PCR tubes
were difficult to work with because of the size and it was hard to label it.
On the third day we made the gels to put our PCR in. The gel was opaque and it
looked similar to rosin. The spaces we had to put our PCR in was very small and it
Analysis:
Our class did not have enough successful results so we made theoretical data to
learn about genotype frequencies. We said there was 15 people with +/+, 10 with +/-,
and 12 with -/-. To find the frequencies you have to find how many positive or negative
alleles and divide by the total number of alleles. For the double positive and double
negative alleles, you take the number you just found and square it. For the +/- you take
the first -/- number and the first +/+ number and you multiply them together and them
multiple that be 2. The number you get is the amount of people who have that set and it
is okay if it is different from the original number. After doing those calculations with the
numbers we were given we got 11 people with +/+, 18 people with +/-, and 9 people
with -/-.
My hypothesis was not correct because I did not get any results. I feel that if I
I think my one of my errors was that I did not rinse the salt rinse for a long
enough amount of time. Also I could have probably been more precise with the pipet
measurements. Next time I can also read through all the directions more closely so I
We can improve this lab by making sure we are all quiet when directions are
given to us so we all hear whats going on. We can also try to manage our time better
so we are not trying to rush in the end of class. This lab could also be better if we had
more results so we can see our own results, but that can not be changed by changing
the procedure.
Conclusion:
I discovered that lab work is very precise. In this lab, we learned about PCR and
gel electrophoresis using our DNA. To do this we had to add a primer mix and a master
mix to our DNA, while heating and cooling it to exact temperatures for certain amout of
time, and b placing it into gels. One part that we had to be extremely exact on was the
time and temperatures of our PCR. If the mix was too hot or too cold, our DNA would
not react in the gels. Making sure that the temperatures and times were accurate was
crucial to the lab if you wanted to see any results. Another part of the lab where we had
to be exact was when we were using the pipet to transfer different liquids from one
place to another. The importance of that is that the lab will only work if you have the
same exact mixture of what has already been tested and proven to work. Any other
mixture will cause error because different amounts of those liquids will have diverse
reactions affecting the PCRs reaction with the gel. Now in the future I know how careful