reverse transcription PCR (DDRT-PCR), introduced firstly by Liang and Pardee (Liang and Pardee, 1992), can identify the total number of genes subject to up- or down-regulation in a particular setting and has been widely used to screen for genes that are differentially expressed (Breyne and Zabeau, 2001; Ahmed et al.
The use of longer primers on differential display
strategies may also lead to detection of DNA polymorphism through RAPD-like results.
The Differential display
was used to detect the fragments of gene/DNA sequences, expressed differentially under stress conditions.
of serum proteins before and after the TCC exercise program.
Study of mRNA expression of Arabidopsis thaliana under stimulation using modified differential display
RT-PCR with silver staining, Colloids Surfaces B: Biointerfaces, 40: 31-34.
This discrepancy between experimental methods is probably due to the limitations of DDRT-PCR, such as the somewhat indiscriminant amplification of a population of similarly sized cDNAs and the difficulty in excising a single amplification product from the differential display
gel (Debouck, 1995).
The original differential display
method relies on the use of an oligo-dT anchord primer and a random primer to amplify the cDNA pools obtained from the control and treated samples in order to allow easy identification and recovery of cDNAs that exhibit differential expression .
Genes differentially expressed in liver and adipose tissue of growth hormone transgenic coho salmon, compared to wild-type controls, were identified using differential display
(GeneFishing DEG System; SeeGene).
Other groups are using bioinformatics such as digital differential display
and in silico Northern blotting to compare gene expression between healthy and cancerous tissues to identify overexpressed genes (3).
Combining the technique of RNA fingerprinting and differential display
to obtain differentially expressed mRNA.
These services include genomic data management, microarray support, quantitative PCR, differential display
, genotyping, genetically-modified organism testing, and a battery of other molecular biology services which support researchers in the pharmaceutical, biotechnology and academic sectors in their attempt to understand the mechanisms of disease and to discover and develop new drugs.
PCR was performed in a 25 [micro]l reaction mixture including lug total RNA, 1 [micro]l of 1 [micro]M primer H-T11A (5'-AAGCTTTTTTTTTTTA-3'), 200U of MMLV reverse transcriptase, 2 [micro]l of 10mM dNTPs and 4 [micro]l of 5X first strand buffer [250 mM Tris (pH 8.