About Differential Display PCR
All living organisms have thousands to tens of thousands of unique genes
encoded in their genome, of which only a small fraction, perhaps 15%, are
expressed in any individual cell. Therefore, it is the temporal and
spatial regulation in gene expression that determines life processes. The
course of normal cellular development as well as pathological changes
that arise in diseases such as cancer are all believed to be driven by
changes in gene expression. A pressing problem is to identify and
characterize those genes that are differentially expressed in order to
understand the molecular nature of disease state and subsequently, to
devise rational therapies. Differential Display was invented in 1992 by
Drs. Arthur Pardee and Peng Liang to allow rapid, accurate and sensitive
detection of altered gene expression (Science. 1992, 257:967; U.S. Patent
5,262,311).
Further readings about differential display technique
- What's Differential Display (GenHunter)
Introduction to differential display technique
- Differential Display (Chun-Ming Liu)
The following procedures are described:
- RNA extraction and qualification
- DNase treatment of RNA sample
- Reverse transcription
- PCR amplification
- Separation on acrylamide gels
- DNA extraction from bands of interest
- Re-amplification by PCR
- Separation on agarose gels
- Excision of amplified products
- Differential Display (Breeden Lab)
Detailed protocol for differential display
- Differential Display (Plant Molecular Biolgy Lab)
It's for plant RNA display. The protocol should be general.
- Differential Display-Reverse Transcription-PCR (Gerard Lazo)
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