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Wednesday, September 10, 2008

Differential Display PCR

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)

Rational primer design greatly improves differential display-PCR.

Applications of Differential-Display Reverse Transcription-PCR.

Differential Display-PCR -- Sambrook and Russell.

Development and optimization of a fluorescent differential display.

The Science Advisory Board – Differential Display PCR.

Perspective: Micoarrays and Differential Display PCR.

Laboratory Techniques: Differential Display - Polymerase Chain Reaction
Effect of Primer Purity on the Banding Patterns of DifferentialDisplay Polymerase Chain ReactionDifferential Display of RNAs (Breeden Lab.)

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