Home

If we knew what it was we were doing, it would not be called research, would it?
Albert Einstein


Quantitative and Qualitative Analysis of Gene Expression

Experimental Molecular Biology SPRING 2016


Lab begins promptly at 1 p.m.; we meet in ABL B03 (same place as for BIOC 311).  You must come to lab prepared--this requires you to read over and think about the experimental protocols, not just print a copy of them. The procedures for each day are available here and you will be given any additional information in the pre-lab lectures.

SPECIAL NOTE: Record enough procedure details in your notebook during each day of lab so that you can repeat these procedures using your notebook as the ONLY resource (i.e., you should not use printed web pages or handouts from previous labs).

Catalogues from biological suppliers represent an accessible and free source of valuable information on reagents, equipment, and technical procedures. We order primarily from these companies:

General Reference:
Sambrook, J. and Russell, D.W. (2001) Molecular Cloning: A Laboratory Manual, Third Edition (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York).


Day 1 Primer design: Tues., 8 March

Discussion Topics

Experimental Procedures

Course introduction

Choosing a reference gene

Overview of research project

Choosing a gene of interest

Eleven Golden Rules of qPCR

Primer design

Primer design

Day 2 Validation of primers for qPCR: Thurs., 10 March

Discussion Topics

Experimental Procedures

Validation of primers

Quantitative PCR (qPCR): temp. gradient

Day 3 Isolation of RNA: Tues., 15 March

Discussion Topics

Experimental Procedures

Isolation of RNA

RNA isolation

Day 4 cDNA synthesis from total RNA: Thurs., 17 March

Discussion Topics

Experimental Procedures

Data analysis (temp. gradient qPCR)

Agarose gel analysis of RNA

Test qPCR set-up

Reverse transcription of RNA

Day 5 Analysis of gene expression with qPCR: Tues., 22 March

Discussion Topics

Experimental Procedures

Data analysis (test qPCR)

Test qPCR

Preparing a Scientific Poster

Poster critique

Day 6 Analysis of qPCR data: Tues., 24 March

Discussion Topics

Experimental Procedures

Data analysis (test qPCR)

Data analysis

Day 7 Analysis of qPCR data: Tues., 29 March

Discussion Topics

Experimental Procedures

Data analysis

Agarose gel of qPCR products

Eleven Golden Rules of qPCR

Data analysis

Day 8 Analysis of qPCR data: Thurs., 7 April

Discussion Topics

Experimental Procedures

Data analysis

Data analysis

*Final notebook pages, including conclusions and Table of Contents, are due by 4 p.m. on Tues., 12 April (please staple pages and put in Dr. Beason's inbox outside ABL 326)
**Posters (PPT file) are due by 11:59 p.m. on Mon, 18 April (submitted in Canvas)
***Poster Q/A session is Thurs., 21 April, in GRB W100D

Copyright, Acknowledgements, and Intended Use
Created by B. Beason (bbeason@rice.edu), Rice University, 21 July 2003
Updated 25 February 2016