Finals-Homework (MOLBIO)

Week 13-14 Polymerase Chain Reaction & Gel Electrophoresis
Please answer the following as brief as possible and in your own words (15 points).
1. Differentiate PCR, qPCR (quantitative PCR) and RT-PCR (Reverse transcription polymerase chain
reaction).
The presence or absence of DNA can be determined using the very straightforward PCR
method. Small fragments of DNA are regularly amplified for sequencing or other
applications down the road. The way that PCR works is that it amplifies genetic material by
making a lot of copies of it, much like a photocopy, to make it easier to detect. Quantitative
polymerase chain reaction (qPCR) is an improved version of PCR that copies more than just
the genetic information, the letter q stands for quantifying. Along with amplifying the
genetic sequence, it also counts how many copies of that sequence are present in the
sample. Reverse transcriptase-mediated polymerase chain reaction (RT-PCR) is a technique
that transforms RNA molecules into their complementary DNA (cDNA) sequences, which are
then amplified using conventional PCR techniques. Purified RNA is used for reverse
transcription.
2. List 5 applications of PCR (5 points).
a. Paternity Testing. For establishing DNA paternity, the PCR approach has become the industry
standard of practice. A typical DNA paternity test today uses buccal swab specimens, which are taken
from each tested party in a non-invasive manner, making it suitable for even newborn children.
b.Sequencing. By specifying the sequence of two oligonucleotides, the polymerase chain reaction can
enzymatically amplify particular DNA regions up to a million times. The development of sequencing
templates, either from cloned inserts or directly from genomic DNA, is one of the method's most
important applications.
c. Cloning. The rapid method of PCR cloning, which is frequently used for activities requiring a higher
throughput than is practical with conventional cloning procedures, allows for the quick replication of
genes. It makes it possible to clone DNA segments that aren't widely accessible.
d. Modification of DNA fragments. Using short synthetic DNA fragments known as primers, PCR selects
a region of the genome to be amplified. This section is then amplified through a number of rounds of
DNA synthesis.
e. Forensics. In just a few hours, the PCR method can reproduce a specific DNA sequence millions of
times. This is crucial for forensic DNA samples because the DNA normally recovered at crime scenes is
scarce and of low quality.
Week 15 Molecular Cloning
Please answer the following as brief as possible and in your own words.
1. Describe the three ways of how microorganisms acquire antibiotic resistance. (10 points)
a. Mutation. Antibiotic resistance develops in some bacteria during the cell replication process.
Antibiotics are less likely to kill microorganisms with the resistant mutation.
b. Horizontal Gene Transfer. Different bacteria cells exchange resistant genetic material. This can occur
in three ways: transformation, transduction, and conjugation.
c. Microorganisms use enzymes and proteins to alter or destroy antibiotics.
2. Give at least 5 applications of molecular cloning in medicine, clinical microbiology and/or industry. (10
points)
a. Creating disease-modeling animal clones. Genetically engineered animals serve as disease-cause

gene mutation models. These transgenic creatures need to be produced via a protracted process of
breeding spanning several generations and trial and error. Cloning might expedite the process of
developing transgenic animal models and offer a population of study animals that are genetically similar.
b. Using cloning to create stem cells. Some researchers are looking into using cloning to produce stem
cells that are genetically similar to a certain person. 2013 saw the first successful cloning of human
embryonic stem cells by Oregon Health and Science University researchers.
c. Recombinant vaccines. The development of recombinant antigens and vaccines depends increasingly
on the science of molecular cloning. Cloning and antigen expression allows for the large-scale
production of antigens that were previously inaccessible due to a shortage of reagents.
d. Antibacterial peptides. Due to the rapidly expanding antimicrobial resistance, increased focus has
been paid to the creation of more powerful antimicrobial peptides that quickly destroy target cells and
are effective against infections that are clinically relevant and antibiotic-resistant. The molecular cloning
of genes that produce antibiotics is actively being registered by many firms.
e. Recombinant cytokines. In several important homeostatic processes, the immunomodulator
cytokines have a crucial role. The genes that encode these proteins have been cloned thanks to
advancements in recombinant DNA technology, enabling researchers to utilize as many of these
cytokines as they like to cure illnesses.
Week 16 Phylogenetics
Please answer the following as brief as possible and in your own words.
1. Give at least 5 significances of phylogenetics (10 points).
a. The phylogenetic tree can be used to trace the evolutionary histories of infectious bacteria.
b. It's utilized to look at the common ancestors of both living and extinct species.
c. Phylogenetics not only teaches us how the sequences came to be the way they are today, but it also
teaches us basic principles that allow us to forecast how they will change in the future.
d. Its objective is to demonstrate evolutionary connections between organisms assumed to have shared
a common ancestor.
e. Its aim is to link the evolutionary milestones of significant life forms to the tree of life.
2. Please explain briefly. Based on the paper Phylogenetic Supertree Reveals Detailed Evolution of
SARS-COV-2, how would you apply phylogenetics in explaining the evolution of COVID-19? (10 points).
 I'll start by identifying or gathering every strain; once we've done that, we can then figure out
how each strain is related to the others. We will be able to identify it and identify the
subfamilies for them. We can use the digital approach to quickly locate or track the
development of COVID-19. We can also predict upcoming strains using phylogenetic analysis.
Week 17 Antibiotic Resistance Genes
Please discuss the following as brief as possible (20 points).
1. Discuss and elaborate what will happen if bacteria become resistant to all antibiotics (10 points).
 When bacteria and fungi learn to resist the drugs meant to kill them, it is said that there is an
antimicrobial resistance problem. The germs persist and develop in light of this. It is typically
more difficult and expensive to treat infections when bacteria become resistant to antibiotics.
Public health is gravely in danger if the capacity to treat deadly bacterial infections is lost.
Diseases can spread, worsen, and, at their worst, kill a patient or other community members.
2. Please enumerate 5 ways in order to help prevent the spread of antibiotic resistant bacteria and/or
antibiotic resistance genes (10 points).
a. Antibiotics should only be used if prescribed by a licensed health professional.

b. Antibiotics should never be shared or used if they are no longer required.
c. Discuss the proper use of antibiotics, antibiotic resistance, and the implications of misuse with the
patients.
d. Facilitate the adoption of a strong national antibiotic resistance action plan.
e. Invest in the discovery and creation of new treatments, vaccines, diagnostics, and other equipment.
Bonus Question (20 points)
What have you learned in Molecular Biology this semester. Give at least 5 things in 5 sentences.
a. The first thing I learned about this subject was the fundamental knowledge a MedTech must possess,
such as the aseptic technique, which is utilized to keep the laboratory workplace clean and prevent
contamination of the tools and samples you are using.
b. The most crucial understanding is laboratory safety because this is where everything begins. If a
laboratory user lacks this information, he or she runs the risk of hurting not only the samples or items in
the lab but also themselves.
c. The streak plate technique is the next, and it seeks to grow isolated colonies of an organism on an
agar plate.
d. I discovered the value of PCR in disease identification, particularly in regard to the current pandemic,
because it is required for an antigen test that amplifies viral RNA collected from patient samples.
e. The central dogma concept, which is related to understanding how genetic information is transferred
from DNA to RNA and finally to proteins, is the last but most important.

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Week 13-14 Polymerase Chain Reaction & Gel Electrophoresis

2. List 5 applications of PCR (5 points).

Week 15 Molecular Cloning

a. Creating disease-modeling animal clones. Genetically engineered animals serve as disease-cause

1. Give at least 5 significances of phylogenetics (10 points).

Please discuss the following as brief as possible (20 points).

a. Antibiotics should only be used if prescribed by a licensed health professional.

Bonus Question (20 points)

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