Biotechnology

Quarter 4 – Module 3:
Steps in Recombinant DNA
Technology

DIVISION OF ANGELES CITY

Biotechnology– Grade 8
Alternative Delivery Mode
Quarter 3 – Module 3: Steps in Recombinant DNA Technology
First Edition, 2021

Republic Act 8293, section 176 states that: No copyright shall subsist in any work of the
Government of the Philippines. However, prior approval of the government agency or office
wherein the work is created shall be necessary for exploitation of such work for profit. Such
agency or office may, among other things, impose as a condition the payment of royalties.

Borrowed materials (i.e., songs, stories, poems, pictures, photos, brand names, trademarks,
etc.) included in this module are owned by their respective copyright holders. Every effort
has been exerted to locate and seek permission to use these materials from their respective
copyright owners. The publisher and authors do not represent nor claim ownership over
them.

Published by the Department of Education

Regional Director: May B Eclar PhD, CESE III
Assistant Regional Director: Rhoda T. Razon EdD, CESO V

Development Team of the Module

Writers: Larissa Manalili

Editors: Sherilyne L. Reyes, Jennifer Praza, Edgardo D. Cortez,
Jenny S. Tongol, Edythe Hipolito
Reviewers: Gemima A. Estrabillo, Emily F. Sarmiento, Hermes Vargas, Adrian
Tamayo, Krislene Ida N. Mercado, Noel S. Reganit
Mercedes Bactol, Billy Ray B. Manuel, Marvin R. Leano, Gemmarie
G. Rivas
Illustrator: Arnold Arceo
Layout Artist: Maricon H. Rivera, Noel S. Reganit
Management Team: May B. Eclar PhD, CESO V
Rhoda T. Razon EdD, CESO V
Ma. Irelyn P. Tamayo PhD, CESE
Fernandina P. Otchengco, PhD, CESE
Librada M. Rubio PhD
Ma. Editha R. Caparas EdD
Rochella C. David
Emily F. Sarmiento PhD
Gemima A. Estrabillo EdD

Printed in the Philippines by ________________________

Department of Education –Region III – Schools Division of Angeles City

Office Address: Jesus St. Pulungbulu, Angeles City

Telefax: (045) 322-5722;322-472 888-0582;887-6099
E-mail Address: [Link]@[Link]
 9
Biotechnology
Quarter 4 – Module 3:
Steps in Recombinant DNA
Technology
Introductory Message

This Self-Learning Module (SLM) is prepared so that you, our dear learners,
can continue your studies and learn while at home. Activities, questions,
directions, exercises, and discussions are carefully stated for you to understand
each lesson.

Each SLM is composed of different parts. Each part shall guide you step-
bystep as you discover and understand the lesson prepared for you.

Pre-tests are provided to measure your prior knowledge on lessons in each

SLM. This will tell you if you need to proceed on completing this module or if you
need to ask your facilitator or your teacher’s assistance for better understanding of
the lesson. At the end of each module, you need to answer the post-test to self-
check your learning. Answer keys are provided for each activity and test. We trust
that you will be honest in using these.

In addition to the material in the main text, Notes to the Teacher are also
provided to our facilitators and parents for strategies and reminders on how they
can best help you on your home-based learning.

Please use this module with care. Do not put unnecessary marks on any
part of this SLM. Use a separate sheet of paper in answering the exercises and
tests. And read the instructions carefully before performing each task.

If you have any questions in using this SLM or any difficulty in answering
the tasks in this module, do not hesitate to consult your teacher or facilitator.

Thank you.

2
 What I Need to Know

This module was designed and written with you in mind. It is here to help
you master the steps in Recombinant DNA Technology. The scope of this module
permits it to be used in many different learning situations. The language used
recognizes the diverse vocabulary level of students. The lessons are arranged to
follow the standard sequence of the course. But the order in which you read them
can be changed to correspond with the textbook you are now using.

The module is about:

● Steps in Recombinant DNA Technology

After going through this module, you are expected to:

1. Outline the steps in Recombinant DNA Technology

3
 What I Know

Directions: Read each question carefully. Choose the letter of the best answer.

1. Which refers to the combination of two DNA strands that are constructed
artificially?
a. Genetic Material
b. Recombinant Cells
c. Recombinant DNA
d. Restriction Enzymes

2. Which among the following is the Blueprint of Life?

a. Cell
b. DNA
c. Protein
d. RNA

3. Which refers to the small accessory ring of the DNA in some bacteria? a.
Interferon
b. Plasmid
c. Restriction enzymes
d. Vector

4. What molecule is being used to cut a specific area of the DNA?

a. Agarose Gel
b. Plasmid
c. Recombinant Cells
d. Restriction enzymes

5. An organism that contains genetic material from two different organisms is

called ___________. a. Clone
b. GMO
c. Mutant
d. Restriction enzymes

6. What technique is being used to make a million copies of a sample DNA?

a. DNA Fingerprinting
b. Gel Electrophoresis
c. Gene Therapy
d. Polymerase Chain Reaction

4
7. Which among the following does NOT describe the principle of Gel
Electrophoresis?
a. DNA fragments are separated on the basis of size.
b. The DNA fragments will move towards the negative charge.
c. The smallest DNA fragments will move faster than the larger DNA
fragments.
d. DNA fragments are injected into wells and an electric current is applied
along with the gel.

8. What equipment is used to amplify the DNA?

a. Centrifuge Machine
b. Microscope
c. Pipette
d. Thermal Cycler

9. Which among the following serves as a starting point for DNA synthesis?
a. DNA polymerase
b. Primer
c. Restriction enzymes
d. Taq polymerase

10. What organism is being used to transfer foreign genetic material into a
cell? a. DNA
b. Plasmid
c. Restriction enzymes
d. Vector

5
 Lesson
Steps in Recombinant DNA
1 Technology
What’s In

Activity 1
Direction: Write True if the statement is correct and False if incorrect.

1. Genetic Engineering is the artificial manipulation of DNA or other nucleic

acid molecules in order to modify an organism or population of organisms.
2. Restriction enzyme was discovered by Werner Arber.
3. Genes are small rings of DNA.
4. DNA technology makes it possible to clone genes for basic research and
commercial applications.
5. Restriction enzymes cut the DNA into a particular site.
In our previous lesson, we learned how genetic materials are manipulated.
One of the ways is through Genetic engineering. In Genetic engineering, genes are
manipulated for practical purposes.

Genetic Engineering

DNA technology has launched a revolution in Biotechnology. DNA technology

(via gene manipulation) makes it possible to clone genes for basic research and
commercial applications. DNA technology is applied to areas ranging from
agriculture to criminal law. One example of DNA technology is Genetic engineering.
Genetic Engineering is the the artificial manipulation, modification, and
recombination of DNA or other nucleic acid molecules in order to modify an
organism or population of organisms. In the latter part of the 20th century,
however, the term came to refer more specifically to methods of recombinant DNA
technology, in which DNA molecules from two or more sources are combined either
within cells or in vitro and are then inserted into host organisms in which they are
able to propagate.

6
 The possibility for recombinant DNA technology emerged with the discovery of
restriction enzymes in 1968 by Swiss microbiologist Werner Arber.

Most recombinant DNA technology involves the insertion of foreign genes into
the plasmids of common laboratory strains of bacteria. Plasmids are small rings of
DNA; they are not part of the bacterium’s chromosome. Nonetheless, they are
capable of directing protein synthesis, and, like chromosomal DNA, they are
reproduced and passed on to the bacterium’s progeny. Thus, by incorporating
foreign DNA (for example, a mammalian gene) into a bacterium, researchers can
obtain an almost limitless number of copies of the inserted gene. Furthermore, if
the inserted gene is operative (if it directs protein synthesis), the modified bacteria
will produce the protein specified by the foreign DNA. Editors of Encyclopedia
Britannica (2020).

What’s New

Activity 1. Steps in Recombinant DNA Technology

Directions: Arrange the steps in Recombinant DNA Technology in chronological
order. Use numbers 1-7.
A. Amplification Using PCR
B. Isolation of Recombinant Cell
C. Isolation of Genetic Material
D. Obtaining or culturing the Foreign Gene product.
E. Ligation of DNA Molecules.
F. Restriction Enzymes Digestion
G. Insertion of Recombinant DNA into Host

What is It

In this lesson, we shall outline the main steps in Recombinant DNA

Technology and the importance of Recombinant DNA Technology.

7
Recombinant DNA Technology

Recombinant DNA technology refers to the joining together of DNA molecules

from two different species that are inserted into a host organism to produce new
genetic combinations that are of value to science, medicine, agriculture, and
industry. Recombinant DNA (rDNA), on the other hand, is the general name for a
piece of DNA that has been created by the combination of at least two different DNA
strands. They are DNA molecules formed by laboratory methods of genetic
recombination to bring together genetic material from multiple sources, creating
sequences that would not otherwise be found in the genome. Aryal (2018).

Steps of Genetic Recombination Technology

1. Isolation of Genetic Material - Since DNA exists within the cell membrane
along with other macromolecules such as RNA, polysaccharides, proteins, and
lipids, it must be separated and purified which involves enzymes such as
restriction enzymes.

2. Restriction Enzymes Digestion - The technique ‘Agarose Gel Electrophoresis’

reveals the progress of the restriction enzyme digestion. This technique involves
running out the DNA on an agarose gel. On the application of current, the
negatively charged DNA travels to the positive electrode and is separated out
based on size. This allows separating and cutting out the digested DNA
fragments. The vector DNA is also processed using the same procedure.

3. Amplification Using PCR - Polymerase Chain Reaction or PCR is a method of

making multiple copies of a DNA sequence using the enzyme – DNA polymerase
in vitro. It helps to amplify a single copy or a few copies of DNA into thousands
to millions of copies. PCR reactions are run on thermal cyclers using the
following components: 1.)Template – DNA to be amplified 2.)Primers – small,
chemically synthesized oligonucleotides that are complementary to a region of
the DNA. 3.) Enzyme – DNA polymerase 4.)Nucleotides – needed to extend the
primers by the enzyme. 5.) The cut fragments of DNA can be amplified using
PCR and then ligated with the cut vector.

4. Ligation of DNA Molecules – The purified DNA and the vector of interest are
cut with the same restriction enzyme. This gives us the cut fragment of DNA
and the cut vector that is now open. The process of joining these two pieces
together using the enzyme DNA ligase is ligation. The resulting DNA molecule
is a hybrid of two DNA molecules – the interest molecule and the vector. In the
terminology of genetics this intermixing of different DNA strands is called
recombination. Hence, this new hybrid DNA molecule is also called a
recombinant DNA molecule and the technology is referred to as the
recombinant DNA technology.

8
5. Insertion of Recombinant DNA into Host - In this step, the recombinant DNA
is introduced into a recipient host cell mostly, a bacterial cell. This process is
called transformation. Bacterial cells do not accept foreign DNA easily.
Therefore, they are treated to make them competent to accept new DNA. The
processes used may be thermal shock, Ca++ ion treatment, and electroporation.

6. Isolation of Recombinant Cells-The transformation process generates a

mixed population of transformed and non-transformed host cells. The selection
process involves filtering the transformed host cells only. For isolation of
recombinant cells from non-recombinant cells, a marker gene of the plasmid
vector is employed.
7. Obtaining or culturing the Foreign Gene product - When you insert a piece
of alien DNA into a cloning vector and transfer it into a bacterial cell, the alien
DNA gets multiplied. The ultimate aim is to produce a desirable protein
expression. The cells harboring cloned genes of interest are grown on a small
scale in the laboratory. These cell cultures are used for extracting the desired
protein using various separation techniques.

Recombinant Human Growth Hormone

Source: [Link]
[Link]

Importance of Recombinant DNA Technology

9
 Recombinant DNA technology is playing a vital role in improving health
conditions by developing new vaccines and pharmaceuticals. The treatment
strategies are also improved by developing diagnostic kits, monitoring devices, and
new therapeutic approaches. Synthesis of synthetic human insulin and
erythropoietin by genetically modified bacteria and the production of new types of
experimental mutant mice for research purposes are some one of the leading
examples of genetic engineering in health. Likewise, genetic engineering strategies
have been employed to tackle environmental issues such as converting wastes into
biofuels and bioethanol, cleaning the oil spills, carbon, and other toxic wastes, and
detecting arsenic and other contaminants in drinking water. The genetically
modified microbes are also effectively used in biomining and bioremediation.

What’s More

Activity 1. Enzymes in Recombinant DNA Technology

Directions: Draw a Venn Diagram to differentiate DNA Polymerase with DNA Ligase.

DNA Polymerase DNA Ligase

Guide Questions
1. What is the first step in Recombinant DNA Technology?
2. In what step does the cut fragment of DNA and the cut vector are joined
together?

10
 3. What is the final step in Recombinant DNA Technology?

Activity 2. Modeling Bacteria Transformation

Directions: Using the word choices provided in the boxes, fill in the numbered
boxes with the steps of bacteria transformation and the lettered lines with the
name of the structure next to them.

Source:[Link]
ctivity1.
Word Choices for Letters Word Choices for Numbers
foreign DNA with the desired bacteria transformed with recombinant
gene plasmid
plasmid recombinant plasmid cut with restriction enzyme
DNA DNA ligase joins sticky ends to form
recombinant plasmid

Guide Questions
1. What is the role of restriction enzymes in Recombinant DNA Technology?
2. What is the function of the DNA Ligase?

11
 3. What is a Plasmid?

Activity 3 . Recombinant DNA Technology

Directions: Read the choices from each numbered item in Column A and identify
which is NOT included from these groups. Then classify it by choosing the correct
answer in Column B.
Note: To get one (1) point from this activity, two (2) responses must be answered
correctly.
Column A Column B
1. a. Primers A. Isolation of Genetic Material
b. DNA Polymerase
c. Agarose Gel B. Restriction Enzymes
d. PCR Digestion

2. a. Transformation
b. Ligation C. Amplification Using PCR
c. Recombinant DNA
d. Ligase D. Ligation of DNA Molecules

3. a. Gel Electrophoresis
b. Protein expression E. Insertion of Recombinant
c. Positive electrode DNA into Host
d. Agarose Gel
F. Isolation of Recombinant
4. a. Marker gene is employed. Cells
b. Filtering of the transformed host cell.
c. Negatively charged DNA travels to the G. Obtaining or culturing the
positive electrode. Foreign Gene product
d. Mixed population of transformed and
non-transformed cells.

5. a. DNA is separated based on size.

b. Cutting out of digested DNA
fragments.
c. Negatively charged DNA travels to the
positive electrode.
d. Amplify a single copy of DNA into
thousands or millions

Guide Questions
1. What are Primers?
2. What is the function of the PCR in Recombinant DNA Technology?

12
 3. What is the charge of the DNA?

Activity 4. Complete the Steps in Recombinant DNA Technology

Directions: Complete the figure below by supplying the missing Step in
Recombinant DNA Technology.
1. Isolation of Genetic Material.

2.

3. Amplification Using PCR.

4.

5.

6. Isolation of Recombinant Cells

7.

Guide Questions
1. What enzyme is used in DNA Ligation?
2. In what step in Recombinant DNA does Transformation occur?
3. What is the function of the Gel Electrophoresis in Recombinant DNA
Technology?

13
Activity 5. Describing the steps in Recombinant DNA Technology
Directions: Identify which step in Recombinant DNA Technology is involved.
Event Steps in Recombinant DNA
Technology
1. Running out the DNA on an agarose
gel.
2. Harboring cloned genes of interest
are grown on small a scale in the
laboratory.
3. DNA must be separated and purified.
4. The recombinant DNA is introduced
into a recipient host cell
5. Making multiple copies of a DNA
sequence
Guide Questions
1. What equipment makes multiple copies of the DNA?
2. How do DNA fragments separate in Gel Electrophoresis?
3. What enzyme is being used to separate and purify the DNA from the cell?

Activity 6. Application of Recombinant DNA Technology

Directions: Read and understand the situation below about the Humulin R. Answer
the questions below after reading the article.

Eli Lilly began producing insulin from animal pancreas but fell short of the
demand, and the potency varied up to 25% per lot. The development of an
isoelectric precipitation method led to purer and more potent animal insulin,
decreasing the variation between lots to 10%. These discoveries led to the
introduction of longeracting animal insulins in the market. Protamine zinc insulin
lasted 24–36 hours. Isophane neutral protamine Hagedorn lasted 24 hours and
could be mixed with regular insulin. The pharmacokinetics and effects of
amorphous lente insulin (semilente, lente, and ultralente) depended on the
proportion of zinc. In 1978, the first recombinant DNA human insulin was prepared
by David Goeddel and his colleagues (of Genentech) by utilizing and combining the
insulin A- and B- chains expressed in Escherichia coli. Thereafter, Genentech and
Lilly signed an agreement to commercialize rDNA insulin. In 1982, the first insulin
utilizing rDNA technology, Humulin® R (rapid) and N (NPH, intermediate-acting),
were marketed. Guide Questions
1. Are you in favor of using the animal pancreas to replace the insulin in the
human body? Why?
2. Based on the article, how does Recombinant DNA benefit humans?
3. What is Recombinant DNA Technology?
4. What is the function of a vector?
5. What are the common vectors in Recombinant DNA Technology?

14
 What I Have Learned

Directions: Fill in the blanks to complete the statements.

1. The general name for a piece of DNA that has been created by the
combination of at least two strands of DNA is called _____________.
2. The process of introducing recombinant DNA into a recipient host cell is
called _____________ .
3. Agarose Gel Electrophoresis involves running out the DNA on an
_____________.
4. The process of joining the cut fragment of DNA and the cut vector together
using an enzyme is called_____________ .
5. The _____________ is a method of making multiple copies of a DNA sequence
using an enzyme.
6. Recombinant DNA technology refers to the joining together of _____________
from two different _____________ that are inserted into a host organism to
produce new genetic combinations.
7. Ligation of DNA molecules uses_____________ to cut the vector.
8. An enzyme called _____________ helps to amplify a single copy or a few copies
of DNA into thousands to millions of copies.
9. PCR reactions are run on _____________ to amplify a single copy or a few
copies of DNA.
[Link] small circular molecules which act as carriers for the DNA fragments are
called _____________.

What I Can Do

Activity 1. Recombinant DNA Technology in our Life

Directions: By giving examples, explain the importance of Recombinant DNA
Technology in the given fields.

1. Health
2. Food
3. Environment

15
 Assessment

Directions: Read each statement carefully. Choose the letter of the correct
answer.
1. Which enzyme is being used to amplify the DNA?
a. Endonuclease
b. DNA Helicase
c. DNA Ligase
d. DNA Polymerase

2. The following statements are all TRUE EXCEPT

a. The DNA is located in the nucleus of the cell.
b. Recombinant DNA is a combination of two different strands of DNA.
c. DNA Ligase is used to amplify a single copy or a few copies of DNA.
d. In Agarose Gel Electrophoresis the negatively charged DNA travels to
the positive electrode and is separated out based on size.

For question nos. 3-9, Arrange in order the steps in Recombinant DNA Technology.
Use letters a to g
3. Amplification Using PCR
4. Isolation of Genetic Material
5. Insertion of Recombinant DNA into Host 6. Obtaining or culturing the
Foreign Gene product.
7. Ligation of DNA Molecules.
8. Restriction Enzymes Digestion
9. Isolation of Recombinant Cells

10. Which of the following refers to chemically synthesized oligonucleotides that

are complementary to a region of the DNA?
a. Agarose Gel
b. Primers
c. Restriction Enzymes
d. Vectors

11. Which of the following is TRUE?

a. DNA is positively charged.
b. Ligase is used to isolate the genetic material.
c. Thermal Cycler cuts fragment of DNA and the vector.
d. Gel Electrophoresis separates the DNA Molecule according to size.

For question nos. 12-15. Identify the steps in Recombinant DNA Technology that
are being described in each statement. The choices are as follows:

16
 a. Amplification Using PCR
b. Isolation of Genetic Material
c. Insertion of Recombinant DNA into Host
d. Obtaining or culturing the Foreign Gene product.
e. Ligation of DNA Molecules.
f. Restriction Enzymes Digestion
g. Isolation of Recombinant Cells

12. Marker gene of plasmid vectors is employed.

13. The aim is to produce a desirable protein expression.
14. The DNA molecule is separated and purified using enzymes. 15. The
recombinant DNA is introduced into a recipient host cell.

Additional Activity

Activity [Link] of Recombinant DNA Technology

Explain how recombinant DNA Technology poses a threat to a population or
ecosystem.

17
 Rubric for the Essay
Category 4 3 2 1
Reflective The idea The idea The idea The idea does
Thinking explains explains attempts to not address the
the the demonstrate student’s
student’s own student’s thinking about thinking and/or
thinking and thinking about learning but is learning.
learning his/her own
vague and/or
processes, as learning
unclear about
well as processes.
implications the personal
for future learning
learning. process.
Analysis The idea is an The idea is an The idea The idea does
in-depth analysis of the attempts to not move
analysis of the learning analyze the beyond a
learning experience learning description of
experience, and the value experience but the learning
the value of the value of
of the derived experience.
the derived the learning to
learning to
learning to the student or
self or others, self or others.
others is
and the
vague and/or
enhancement
of the unclear.
student’s
appreciation
for the
discipline.
Making The idea The idea The idea The idea does
Connections articulates articulates attempts to not articulate
multiple connections articulate any connection
connections between connections to other
between this between learning or
this learning this
experiences.
learning experience learning
experience and content experience
and content from past and content
from past learning from past
learning, experiences, learning
life and/or future experiences,
experiences goals. or personal
and/or future goals, but the
goals. connection is
vague and/or
unclear.

18
References
Aryal, Sagar, and Rashid Eltayeb. 2020. “Recombinant DNA Technology- Steps,
Applications and Limitations: Molecular Biology.” Microbe Notes.
[Link]
applicationsand-limitations/.

Bacteria Transformation - Activity.” 2019. [Link]. June 20, 2019.

[Link]
tivity1.

Griffiths AJF, Miller JH, Suzuki DT, et al. 2020. An Introduction to Genetic
Analysis.
7th edition. New York: W. H. Freeman; 2000.
[Link]

Payal. 2016. “7 Main Stages of Recombinant DNA Technology.” Biology Discussion.

[Link]
7main-stages-of-recombinant-dna-technology/56320.

Quianzon, Celeste C., and Issam Cheikh. 2012. "History of insulin." Journal of
community hospital internal medicine perspectives 2, no. 2: 18701.

Rabago, Lilia M, Joaquin, Crescencia C., and Lagunzad, Catherine Genevieve B.

Functional. [Link] biology: modular approach, Vibal Publishing
House, Inc. Metro Manila, Philippines.

The Editors of Encyclopedia Britannica. 2020. “Genetic Engineering.” Encyclopedia

Britannica. Encyclopedia Britannica, Inc.
[Link]

19
For inquiries or feedback, please write or call:

Department of Education – Region III – Schools Division of Angeles City

Jesus St. Pulungbulu, Angeles City, Pampanga, Philippines 2009

Telefax: (045) 322-5722;322-472; 888-0582;887-6099 Telefax:

(632) 8634-1072; 8634-1054; 8631-4985