PCR Lab Write Up

Commented [1]: this needs to cover all of the

Purpose: purposes
The purpose is to list and explain the importance of each component of PCR, to able to
make the PCR happen, and to be able to understand each steps of the procedure.

Hypothesis:
If our predicted allele frequencies are different from our actual allele frequencies then
that means that we are still evolving. Commented [2]: need to be what you think will happen

Procedure:
http://babec.org/wp-content/uploads/2016/12/Alu_Student_Guide_2017.pdf

Material:
http://babec.org/wp-content/uploads/2016/12/Alu_Student_Guide_2017.pdf

Commented [3]: cite

Safety:
http://babec.org/wp-content/uploads/2016/12/Alu_Student_Guide_2017.pdf

Commented [4]: supposed to be observations not

Data/Observation: procedure. need picture of data, label, and
Day 1: We swirled 10 ml of saline solution in our mouth, and spit it out. Then we observations

centrifuge it and saw a small white cluck at the bottom of our microfuge tube. Then we
pour some of the liquids out and placed them on a heating block for 10min. After
heating we centrifuge the tube for 1 min. After the centrifuge, we withdraw 50 microliter
of supernatant from the tube to a newly labeled tube. Then we place the DNA tubes in
a class rack and let our teacher to refrigerate them.

Day 2: We first obtained a tiny PCR tube, and then we pour one after another of 20
microliters of master mix, 20 microliter of primer mix, and 10 microliter of our extracted
DNA into our PCR tube. Then we place them into a thermal cycler. The cycling
protocol for amplification of Alu PV92 is: 1) 95C hold for 2 minutes. 2) 30 cycles of:
94C for 30 seconds 60C for 30 seconds 72C for 2 minutes. 3) 72C hold for 10
minutes. 4) 4C hold, infinity.

Day 3: We retrieved our PCR tube and centrifuge it for 10 seconds. Then we added 5
microliter our loading dye to our PCR tube. Then we carefully loaded 17 microliter of
the loading dye mixture into a well in our gel. Then we loaded 7 microliter of 100 bp
ladder into one of the wells of each gel. Then we hooked our gel box to a power supply
at 150 Volts for 25 minutes. After that our gels are ready to stain and photograph.

Based on the procedure one person got +/-, two person got -/-, one person got +/+.

Analysis/Discussion:
+/- meaning they have Alu insert on one of the chromosomes.
-/- meaning they don't have any Alu inserts on their chromosomes.
+/+ meaning they have Alu inserts on both of their chromosomes. Commented [5]: good but need to say what they look
like on gel

Our hypothesis is not proven because we did not have enough data to support the
hypothesis. Commented [6]: just because no data does not mean
wrong

We could of done many human error include not adding or withdrawing enough of a
certain substance. We could of improved of this lab by simply having people follow the
instruction.

Commented [7]: Follow CLEAR

Conclusion:
From this lab we discovered that we have to follow the procedure precisely or we will
not get any results. This lab is about us understanding the processes of PCR, and
experience these steps for ourselves. It is also about calculate our allele frequencies to Commented [8]: do not restate purpose supposed to
be what you did
see if humans is still evolving. After following the procedure only 4 people got results, 1
person got +/-, 2 people got -/-, and 1 person got +/+. These data was insufficient to
prove anything, it also mean not many people follow the procedure correctly, and next
time we need to strictly follow the procedure.