At the center of the technique is the polymerase chain reaction (PCR) used in this case to amplify sets of message sequences. With PCR, the primers are the key, and for differential display, two sets of primers are most often used: a set of oligo (dT) downstream primers anchored to the poly-A tail of mRNAs, and a set of arbitrary primers, designed to hybridize upstream from the poly-A tail to multiple messages. [The oligo (dT) primers are anchored with one or two bases at the 3' end so that the primer anneals specifically at the 5'-end of the poly-A tail, and does not bind to multiple sites along the tract before getting to the informational part of the message]. The PCR products, which can vary in size from tens of bases to kilobases, are then run out on high-resolution acrylamide gels, from which bands of interest can be isolated and ...
Expression Detection: Identify Differentially Expressed Genes with Differential Display Kits
Differential display, invented in 1992 by Liang and Pardee, ( Science, 257:967-71, 1992) has, in the few years since its description, become the premier technique for studying gene expression. Accordingly, a number of companies have jumped into the market with kits and tools for using this technology. The beauty of this method is that it enables side-by-side comparisons of complex expression patterns from as many samples as can fit on a gel, and eliminates the need for what can be at times tedi
Written byLaura Defrancesco
| 13 min read
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