Research Methods in Genetic Biotechnology W13
Dawn Marshall - SN2102, 864-7677, dawnm-at-mun.ca
Dates: April 22-May 10, 2013
Lectures SN4083 - Laboratory SN3119 - Computer Work SN4117
Last updated: 6May13
Notes, Readings, and Lab Protocols
Module 1 - Course Introduction and Primer Design - Monday 22Apr13
Tentative Schedule (particular days and especially TIMES are subject to change)
Review - the structure of a gene
Module 2 - Lab Safety, Calculations, and Pipetting Skills - Tuesday 23Apr13
Lab Protocol and Calculation Questions
Module 3 - DNA Extraction - Wednesday 24Apr13
Qiagen QIAamp DNA Mini Kit Manual
Reading #3 - DNA and Spectrophometry Reading #4 - Agarose Gel Electrophoresis
Module 4 - RNA Extraction - Thursday 25Apr13
Qiagen RNeasy Fibrous Tissue Mini Kit Manual
Module 5 - cDNA Preparation - Friday 26Apr13
Lecture and Laboratory Protocol
Clontech SMARTer cDNA Synthesis Manual
Qiagen Quantitect cDNA Synthesis Manual
Module 6 - PCR - Monday 29Apr13
Lecture and Laboratory Protocol
QiaQuick PCR Purification Manual
Module 7 - Molecular Cloning - Tuesday 30Apr & Wednesday 1May13
Reading #7 - Plasmid vectors in cloning
Module 8 - Sequencing - May 2-3, 2013
Reading #8
Data Analysis and Final Report May 6-10, 2013
Data Analysis: Sequence data editing
Final Report Information Part I, PartII
Some references:
Aguirre-Hernadez and Sargan 2005
Essay Questions
Q1 . Compare and contrast two different DNA extraction methods, considering such factors as the type of organism/tissue used, how the method works, the cost, scale, and time frame of the study, and the quality/quantity of the isolated DNA.
Q2. Discuss some of the major differences between an RNA-based study and a DNA-based study, and how they are related to differences in the function and structure of the two nucleic acids.
Q3. Compare and contrast two cDNA synthesis methods, in terms of strategy for first and second strand synthesis, usefulness for different downstream procedures, and research question.
Q4. Discuss the role of primer selection in ensuring the specificity of PCR.
Q5. Discuss the role of plasmid vectors in cloning.
Q6. Discuss the modification to Sanger sequencing that enabled both automation and the first human genome sequencing effort.
OR
Compare and contrast Sanger sequencing with the various "next generation" sequencing technologies.
