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Molecular
Biology Protocols Marine
DNA Sequencing and Analysis Center Salsbury
Cove, Maine 04672 USA December 2006 Version |
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1. An Introduction to Primer Design for PCR
2. Purification of Total DNA from Animal Tissues
3A. Purification of Total RNA from Animal
Tissues: Phenol-Based Procedure
3B. Purification of Total RNA from Animal
Tissues: Filter-Based Procedure
4. RNA Analysis with the Agilent 2100
Bioanalyzer
5. Reverse Transcription: SuperScript III
First-Strand Synthesis
6. Polymerase Chain Reaction Amplification of
cDNA Templates
7. Agarose Gel Electrophoresis of PCR Products
8. QiaQuick Gel Extraction Protocol
9. Preparing Gel-Purified PCR Products for
Sequencing
11. Analysis of Gene Expression by Real-Time
Quantitative PCR
12. Microarray Analysis of Gene Expression
This
material is based upon work supported by the National Science Foundation under
Grant No. IBN-9807539 and DBI-0100394. Any opinions,
findings, and conclusions or recommendations expressed in this material are
those of the author and do not necessarily reflect the views of the National
Science Foundation.
Undergraduate
molecular biology courses based on these protocols are supported by the Maine
Biomedical IDeA Network for Biomedical Research
Excellence centered at the Mount Desert Island Biological Laboratory (NCRR NIH
1P20RR16463). The protocols are intended
for teaching purposes and may be freely copied by faculty and students. If used for research, the appropriate citation
for protocols 1-3 and 5-10 is: Towle,
D.W., R.S. Paulsen, D. Weihrauch, M. Kordylewski, C. Salvador, J.-H. Lignot,
and C. Spanings-Pierrot. 2001. Na++K+-ATPase in gills
of the blue crab Callinectes sapidus:
cDNA sequencing and salinity-related expression of α-subunit mRNA and
protein. Journal of Experimental Biology
204: 4005-4012.
These
protocols are online at http://www.mdibl.org/~dtowle/mbw/