Primer design for a prokaryotic differential display RT-PCR

Use of novel downstream primers for differential display RT-PCR.

The polymerase chain reaction (PCR)-based mRNA differential display (3) has been widely used for identifying differentially expressed mRNA in a variety of biological systems (4). mRNA differential display consists of two basic steps: (i) reverse transcription (RT) using a set of 3′-anchored primers, T12MN (M = G, A or C; N = G, A, T or C) and (ii) PCR amplification of cDNA fragments using an ar...

Modification of primer design facilitates the use of differential display.

The method of differential display developed by Liang and Pardee (10) has gained a large audience and is now the method of choice to quickly identify and isolate differentially expressed genes in several experimental systems. The method is powerful because of its inherent simplicity, and it allows the simultaneous comparison of upand down-regulated genes with small amounts of raw material. Neve...

TRIINU KÕRESSAAR Improvement of PCR primer design for detection of prokaryotic species

Differential Display-PCR.

This method uses PCR to amplify and display many cDNAs derived from the mRNAs of a given cell or tissue type. The method relies on two different types of synthetic oligonucleotides: anchored antisense primers and arbitrary sense primers. A typical anchored primer is complementary to approx. 13 nucleotides of the poly(A) tail of mRNA and the adjacent two nucleotides of the transcribed sequence. ...

PCR Primer Design

To make PCR a specific, efficient and cost effective tool for researchers and clinicians the most important aspect is oligonucleotide primer design. This review discusses various aspects of primer design. Advice is provided for optimal design and the role of bioinformatic tools is highlighted. The authors discuss theoretical considerations and compare computational and experimental studies.