Genetics 4013281 Final Lab Exam Fall 2020

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(Q1)
A. One of the most common applications of using recombinant DNA techniques like PCR,
restriction enzyme digests, and agarose gels is to test samples of human cells such as
blood cells to identify people for forensic analysis or paternity testing. This problem is
designed to show you how this type of analysis, called DNA fingerprinting, can be used
to determine paternity. There are three babies (Baby A, Baby B and Baby C) in a
maternity ward, and three sets of confused and worried parents. (Father and Mother #1
are a couple, as are Father and Mother #2, and Father and Mother #3.) This problem will
show you how to figure out which baby goes with each set of parents. As we have spoken
about in class, most of the human genome (>95%) is not genes. Most of the DNA
sequence differences between humans are found in these non-coding regions. Some of
these non-coding regions are just series of DNA sequence repeats found over and over
again. Different humans can have different numbers of repeats at these regions (i.e. “n”
shown below can vary). The way you assay how many repeats someone has is by doing a
PCR reaction using his/her DNA as a template. The primers are designed to the
sequences flanking the repeated region. For instance, take the DNA sequence below.
Say it is found somewhere on chromosome #15. Different humans differ by the value of
“n” (the number of TTAGGAT repeats). You do each PCR reaction and load each one
into a separate well of an agarose gel, and then run the gel.

You obtain the following results.

Genetics 4013281 Final Lab Exam Fall 2020

(a) Why is it that having more repeats leads to a band that is higher in the gel?

Having more repeats lead to longer DNA fragments then it will migrate slowly in gels.

(b) Why is it that some people show only one band?

They have the same number of repeats and In this area, they are homozygote.

(c) Why is it that some people show two bands?

Because They have different numbers of repeats and they are heterozygous in this area

(d) Given the data so far, for which of the three babies (only Baby A, only Baby B, or only Baby
C, all of them, or none of them) can you already conclusively tell who the parents are?

Baby A , and couple #3 because baby A has a mark at n=25 and the only parents who can
given it to the babe is dad#3 from the couple #3.
Genetics 4013281 Final Lab Exam Fall 2020

B. Other parts of the non-coding regions in our genome are not genes but they are also not
regions of repeats. Humans can vary by DNA sequence at these sites, instead of varying
by number of repeats in a row. For instance, take the DNA sequence below. Say it is
found somewhere on chromosome #7. Different humans differ by which basepair is
found at the position marked in bold below; some people have a T-A basepair, whereas
others have an A-T basepair at this bolded position. It just so happens that one version of
this site can be cleaved by a restriction enzyme that recognizes the sequence 5’-
TTGCAA-3’ and cuts between the two Ts. The way you assay which sequence someone
has at this region is by doing a PCR reaction using his/her DNA as a template. The
primers are designed to the sequences flanking the site of variable sequence; each primer
is about 20 nucleotides long. You then treat the PCR product with the restriction enzyme
and run the products of the digestion reaction on an agarose gel.

You obtain the following results.

 Genetics 4013281 Final Lab Exam Fall 2020

(e) Why is it that some people only have one band?

For an allele that can not be cut, they are homozygotes.

(f) Why is it that some people show two bands?

For an allele that can be cut, they are homozygotes.

Why is it that some people show three bands?

They have one allele that can be cut and one allele that can not be cut. They are
heterozygotes.

Match the babies up with their actual parents.

BABY PARENTS

Baby A Father and Mother #1

Baby B Father and Mother #2

Baby C Father and Mother #3

Genetics 4013281 Final Lab Exam Fall 2020

(g)Now go back and look at the first gel drawn in the problem, and answer the question: which
number of repeats did Baby C inherit from his mother?

30 repetitions

(Q2)

As part of an interview for a research position, three applicants are asked to transfer 150 µL of
distilled water with a P-200 micropipette to a weighing paper and to determine the weight of
each drop with an analytical balance. The three measurements made by each of the applicants
are listed below. Use the data below to calculate the mean and standard deviation of the
measements made by each student.

Applicant A: 0.161 g, 0.147 g, 0.142 g

Applicant B: 0.158 g, 0.156 g, 0.157 g
Applicant C: 0.143 g, 0.153 g, 0.150 g

A. Which applicant makes the most precise measurements?

B, is more precise because the measurements are close to each-other.

B. Which applicant makes the most accurate measurements?

C, is more accurate because the mean was 0.148 and the true value was
0.150.
C. Which applicant would you hire? Why?

C and B. Because I l to see good precise and accurate measurements.

Genetics 4013281 Final Lab Exam Fall 2020

(Q3)

A. DNA is soluble in water, but not in isopropanol or ethanol. What does this fact have to do
with our method of extraction? Explain what happened when the isopropanol came in
contact with the plant extract.

In the DNA extraction buffer, the DNA was soluble, but we were not able to see it. It
clumped together as it was stirred into the isopropanol and formed thicker and thicker
strands that were wide enough for us to see

B. In order to study our genes, scientists must first extract the DNA from human tissue.
Would you expect the method of DNA extraction to be the same for human DNA as it
was for plant DNA? Why or why not?

Animal cells do not have cell walls, unlike plant cells. Therefore the cellulose debris from
an animal cell need not be filtered out. If contained in a hypotonic or lower-pressure
solution, animal cells may also be lysed or disintegrated.

C. List two reasons why a scientist might want to study the DNA of human.

1. and appreciate the role of the human genome and how to regulate it; devolve methods
and avoid the disease.

2. More DNA analysis will help solve unresolved crimes by DNA profiling, justice will be
done.

D. Why do we need to quantify DNA?

It is a very important step in many procedures where it is appropriate to know the amount of
DNA that is present while performing restriction digests or performing various techniques
such as PCR and RAPDs for many molecular biological analysis methods and can also
assess their effectiveness.
Genetics 4013281 Final Lab Exam Fall 2020

E. What are the main requirements of GLP (Good Laboratory Practice)?

1. Resources: organization, staff, equipment and services

2. Characterization: objects of test and structures of test
3. Rules: preparations for the analysis (or protocols) and written procedures
4. Outcomes: raw information, final report and archives
5. Assurance of consistency.