Fall 2022 - MBB 308 D100

Molecular Biology Laboratory (3)

Class Number: 2296

Delivery Method: In Person

Overview

  • Course Times + Location:

    Mo 2:30 PM – 4:20 PM
    AQ 3150, Burnaby

  • Instructor:

    Stephanie Vlachos
  • Prerequisites:

    or corequisite: MBB 331 with a minimum grade of C-.

Description

CALENDAR DESCRIPTION:

Modern molecular biological and recombinant nucleic acid methods will be covered. Examples are DNA and RNA isolation, plasmid preparation, restriction enzyme digestion, DNA cloning and polymerase chain reaction. Students with credit for BISC 357 may not take this course for further credit.

COURSE DETAILS:

This course will introduce students to recombinant nucleic acid methods. Lab time will involve one full afternoon every week, for which attendance is mandatory—unexcused absences will be penalized.

Laboratory Exercises:

  1. Check-in, safety, Pipette practice, NanoDrop
  2. Making LB Bacterial plates, casting and loading agarose gels, streaking pJ014 and pJT0 cells onto LB-Amp plates           
  3. Inoculation of liquid culture from pJT0 plates, colony PCR, plasmid miniprep, PCR confirmation via gel electrophoresis    
  4. Restriction digest of PCR product and purified pJT0 plasmid; gel electrophoresis; gel purification of PCR product           
  5. Dephosphorylate of pJT0 vector, ligation, transformation in DH5α cells; Pour LB-Amp     
  6. Site-directed mutagenesis via PCR cycling; DpnI digestion   of parental plasmid and transformation/plating       
  7. Fluorescent characterization of mCherry & mutants
  8. Isolation of genomic DNA and total RNA from Drosophila, NanoDrop.       
  9. Formaldehyde gel analysis, PCR of gene-specific fragment from genomic DNA; generation of a specific cDNA via RT-PCR; pour LB-Amp plates
  10. Gel electrophoresis of the genomic PCR product and cDNA from RT-PCR; spin column purification; cloning of the genomic PCR product into pGEM-T Easy: ligation and transformation   
  11. Miniprep of pGEM-T Easy – genomic PCR, diagnostic restriction digests and gel electrophoresis of cloned product; cycle sequencing of genomic and cDNA clone         
  12. Computer lab for analysis of Drosophila sequencing data
Experiments are subject to change or rearrangement.

Grading

  • Lab Notebook 15%
  • Lab Performance 20%
  • Lab Quizzes 10%
  • Mini-assignment 5%
  • Midterm exam 15%
  • Final exam 20%
  • Lab exam 15%

Materials

MATERIALS + SUPPLIES:

Safety goggles or face shield, and a lab coat are required.

A non-refundable fee of $12.50 will be assessed for the laboratory manual.

RECOMMENDED READING:

Dale, Jeremy W. and Malcolm von Schantz. (2012).  From Genes to Genomes and Applications of DNA Technology, (3rd Ed.). Wiley.
ISBN: 9780470683859

REQUIRED READING NOTES:

Your personalized Course Material list, including digital and physical textbooks, are available through the SFU Bookstore website by simply entering your Computing ID at: shop.sfu.ca/course-materials/my-personalized-course-materials.

Department Undergraduate Notes:


  • For help with writing, learning and study strategies please contact the Student Learning Commons at
    http://learningcommons.sfu.ca/
  • Students requiring accommodations as a result of a disability, must contact the Centre for Accessible Learning (778-782-3112 or e-mail:  caladmin@sfu.ca)

Registrar Notes:

ACADEMIC INTEGRITY: YOUR WORK, YOUR SUCCESS

SFU’s Academic Integrity website http://www.sfu.ca/students/academicintegrity.html is filled with information on what is meant by academic dishonesty, where you can find resources to help with your studies and the consequences of cheating. Check out the site for more information and videos that help explain the issues in plain English.

Each student is responsible for his or her conduct as it affects the university community. Academic dishonesty, in whatever form, is ultimately destructive of the values of the university. Furthermore, it is unfair and discouraging to the majority of students who pursue their studies honestly. Scholarly integrity is required of all members of the university. http://www.sfu.ca/policies/gazette/student/s10-01.html