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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 in Arabidopsis

Experimental Molecular Biology SPRING 2012

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., 6 March

Pre-Lab Topics		Experimental Procedures
Course introduction		Primer design
Overview of research project
Polymerase chain reaction (PCR)
Primer design

Day 2 Isolation of Arabidopsis RNA: Thurs., 8 March

Pre-Lab Topics		Experimental Procedures
Isolation of RNA		RNA isolation
		Agarose gel analysis of RNA

Day 3 cDNA synthesis from total RNA: Tues., 13 March

Pre-Lab Topics		Experimental Procedures
Quantitative PCR (qPCR)		Reverse transcription of RNA

Day 4 Validation of primers for qPCR: Thurs., 15 March

Pre-Lab Topics		Experimental Procedures
Optimization of primers		qPCR

Day 5 Analysis of gene expression with qPCR: Tues., 20 March

Pre-Lab Topics		Experimental Procedures
qPCR optimization		Test qPCR
Preparing a Scientific Poster		Poster critique

Day 6 Analysis of qPCR data: Tues., 27 March

Pre-Lab Topics		Experimental Procedures
Data analysis		Data analysis

Day 7 GUS staining of transgenic Arabidopsis: Thurs., 29 March

Pre-Lab Topics		Experimental Procedures
Reporter		Staining of transgenic plants
GUS staining

Day 8 Observation of GUS-stained plants: Tues., 3 April (meet in ABL 141)

Pre-Lab Topics		Experimental Procedures
Preparing slides		Observation of plants under dissecting scope

*Posters (printed and pdf) are due by 4 p.m. on Tues, 17 April (in ABL 326)
**Poster Q/A session is Thurs., 19 April (in ABL 329 Conference Room)

Copyright, Acknowledgements, and Intended Use
Created by B. Beason (bbeason@rice.edu), Rice University, 21 July 2003
Updated 20 July 2011