BOT103 - Nucleic Acid-Based Technology 1

Outline info
Last revision date 2017-05-29 00:41:16.87
Last review date 2017-07-17 00:16:33.669

Subject Title
Nucleic Acid-Based Technology 1

Subject Description
In the first nucleic acid-based technology course, DNA-based techniques are the focus. In this course, students will acquire both the theoretical and practical knowledge in widely used DNA-based techniques. Examples include sample preparation, extraction, purification and quantification; DNA sample electrophoresis; genetic engineering; southern blot; and PCR (Polymerase Chain Reaction). Teaching will be conducted in both lecture and laboratory formats.

Credit Status
One credit towards the Applied BioChem Techniques (ABCT) Certificate Program (APPLIED BIOTECHNOLOGY).

Learning Outcomes
Upon successful completion of this subject the student will be able to:

  1. Explain the theoretical based of commonly used DNA- based techniques.
  2. Perform DNA isolation, extraction, purification and quantification
  3. Perform various genetic engineering techniques
  4. Perform various PCR tasks
  5. Design complex experiments to achieving goals

Cheating and Plagiarism
Each student should be aware of the College's policy regarding Cheating and Plagiarism. Seneca's Academic Policy will be strictly enforced.

To support academic honesty at Seneca College, all work submitted by students may be reviewed for authenticity and originality, utilizing software tools and third party services. Please visit the Academic Honesty site on for further information regarding cheating and plagiarism policies and procedures.

All students and employees have the right to study and work in an environment that is free from discrimination and/or harassment. Language or activities that defeat this objective violate the College Policy on Discrimination/Harassment and shall not be tolerated. Information and assistance are available from the Student Conduct Office at

Accommodation for Students with Disabilities
The College will provide reasonable accommodation to students with disabilities in order to promote academic success. If you require accommodation, contact the Counselling and Disabilities Services Office at ext. 22900 to initiate the process for documenting, assessing and implementing your individual accommodation needs.

Completion of BOT 100

Topic Outline
Week 1:  tests, exam dates and students’ presentations assignment. Introduction to DNA-based techniques: methods of DNA extraction from various microorganisms, including Gram +,     Gram – bacteria and yeasts. DNA isolation, purification and analysis. Laboratory: Microbial sub-culture and determination of optimal growth conditions for DNA extraction; extraction of genomic DNA from Staphylococcus epidermidis, Escherichia coli and yeast; extraction of plasmid DNA from transformed Escherichia coli.

Week 2: restriction enzyme analysis of DNA: applications; separation and visualization of fragmented DNA by agarose gel electrophoresis; determination of agarose concentration based on DNA size; gel, buffer and electrophoresis reagents preparation; DNA sample preparation and separation; staining and visualization. Laboratory: electrophoretic analysis of microbial DNA; banding pattern analysis of genomic DNA by restriction enzyme electrophoretic analysis.

Week 3: review of DNA recombinant technology: principles and applications. Laboratory: DNA restriction; purification and sub-cloning of a DNA fragment containing a luciferase coding gene; selection and preparation of vector and insert, purification using preparative agarose gel, ligation and transformation of competent cells.

Week 4: principles and mechanisms of bacterial, yeast and mammalian cell transfection. DNA transfection protocols. Laboratory: transfection of a recombinant plasmid containing a luciferase gene into competent E.coli; selection and analysis of transformants by DNA restriction enzyme electrophoretic analysis and luciferase expression.

Week 5: mid-term theory and practical test: identification, selection and characterization of positive recombinant clones from transformed E.coli cultures.

Week 6: Polymerase Chain Reaction (PCR): reverse transcription polymerase chain reaction (RT-PCR) and real-time polymerase chain reaction (qPCR); principle and application. Laboratory: confirmation of successful subcloning and correct gene insertion into recipient plasmid by PCR; primers design and template preparation; PCR amplification and analysis of luciferase genes in selected transformants.

Week 7: Southern blot analysis: principles and applications; Laboratory: Separation of genomic DNA extracts from various microorganisms and from E.coli containing a recombinant plasmid by agarose gel electrophoresis and transfer of fragmented DNA to nitrocellulose/nylon membranes; pre-hybridization and hybridization with  specific DNA probes.

Week 8: DNA probes; application of the principles of nucleic acid hybridization; preparation of DNA template and labeling techniques. Laboratory: detection and analysis; discussion of results.

Week 9: Introduction to genomics; definition; technical and scientific foundation of genomics; recombinant DNA, DNA sequencing methods and bioinformatics; application of genomics to different biotechnology sectors. Students’ presentation.

Week 10: theory and practical final exam.

Mode of Instruction
Lectures, Laboratory, Tutorials, Assignment Presentations, Discussions

Prescribed Texts
to be introduced by the instructor.

Reference Material
instructor’s handouts, technical manuals, laboratory notes, reports

Required Supplies
Laboratory coat, safety glasses

Promotion Policy

Grading Policy
A+ 90%  to  100%
A 80%  to  89%
B+ 75%  to  79%
B 70%  to  74%
C+ 65%  to  69%
C 60%  to  64%
D+ 55%  to  59%
D 50%  to  54%
F 0%    to  49% (Not a Pass)
EXC Excellent
SAT Satisfactory
UNSAT Unsatisfactory

For further information, see a copy of the Academic Policy, available online ( or at Seneca's Registrar's Offices.

Modes of Evaluation

  • Assignments are due at the beginning of the class on which they are due.
  • A late penalty of 10% per day is assessed for late assignments, including those not handed in at the beginning of class when due.
  • Material will not be accepted after one week following the due date and/or when the marked material is returned to students, whichever comes first.
  • Assignments are to be prepared by computer.

Absenteeism and Exams/Quizzes
  • Students should be aware that absenteeism almost guarantees an inability to achieve satisfactory grades.
  • Students who are absent for an examination due to an emergency (e.g., motor vehicle accident, hospitalization or death in the family) may provide official documentation within five days of the missed exam and be provided a deferred exam at a later date.  Official documentation includes a death notice or an original doctor’s certificate identifying the date, length of time expected absence and the specific reason for the absence.  Examinations missed without official documentation and approval result in a grade of zero.
  • There are no deferred options for missed quizzes.  The value of missed quizzes, at the discretion of the Faculty, will be added to other evaluation components.

English Proficiency
  • All written work should demonstrate the following characteristics for clarity and conciseness:
-writing is consistent with the rules of English grammar
-spelling and punctuation are correct
-sentences are structured correctly
-main ideas are supported with specific, relevant examples and reasons
-work flows logically through supporting statements/paragraphs
-work is arranged in correct format (e.g., as a report, essay)
-up to 10% of the final grade may be deducted on all work if the above English competencies are not met.

Format for Assignments
  • Students must use the standard, APA style for quoting sources.   Help is available at:

Laboratory Attendance
The laboratory component is essential and therefore it is strongly recommended that you attend all labs.  Any missed labs must be supported with a legal document with three days of the lab.  Any student who fails to attend more than 2 laboratory classes for a 10 week subject will not pass the subject.    

Laboratory Safety
Students are required to review and understand the safety procedures and guidelines outlined on the first class and sign the sheet to this effect before beginning work in the laboratory.  Students must also wear a lab coat and safety glasses when conducting experiments.
A student who arrives without the proper safety equipment will not be permitted to participant in the lab but will be asked to leave the class.  The student will receive no grade for the lab missed.

Grading is based on the following marking scheme:
Laboratory notebook     10%
Assignments 20%
Theory and practical mid-term test 30%
Theory and practical final exam  40% 


Other Evaluation Considerations
The student is expected to comply with the Safety Rules for working in the laboratory, sign the safety contract, and WILL NOT UNDER ANY CIRCUMSTANCES WEAR CONTACT LENSES in the laboratory. The student will know where all safety equipment is located in the laboratory and will be familiar with WHIMIS concepts and signage.


Approved by: Denis Gravelle