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Differential Display PCR (DDRT-PCR) (CAT#: STEM-MB-0225-WXH)

Introduction

Differential display (also referred to as DDRT-PCR or DD-PCR) is a laboratory technique that allows a researcher to compare and identify changes in gene expression at the mRNA level between two or more eukaryotic cell samples. It was the most commonly used method to compare expression profiles of two eukaryotic cell samples in the 1990s. By 2000, differential display was superseded by DNA microarray approaches.




Principle

The basic approach utilizes different combinations of PCR primers to generate subpopulations of DNA species that are then analyzed on a sequencing gel. Comparison of control and test sample lanes then allows identification of potential differentially modulated mRNAs. In this manner, both increased and decreased levels of individual mRNA species can be determined.

Applications

Compare and identify changes in gene expression at the mRNA level between two or more eukaryotic cell samples.

Procedure

General procedure:
1. Denaturation
2. Annealing
3. Extension