Primers, for polymerase chain reaction

Marks JD, Tristem, Karpas A, Winter G, Oligonucleotide primers for polymerase chain reaction amplification of human immunoglobulin variable genes and design of family-specific oligonucleotide probes, Eur. J. Immunol., 21 985-991, 1991. 

The solid-phase synthesis of DNA oligomers has reached a high level of maturity and is extensively used for the synthesis of primers for polymerase chain reaction or the design of new genes (13). In contrast to the cellular process, the synthetic route forms the polymer in 3 -to-5 direction. The oligomerization is carried out on CPG using a base labile linker. Typically, the nucleobases are introduced as acyl-protected phosphoramidites,... 

Fransen,K Zhong, P., De Beenhouwer, H Carpels, G Peeters, M., Louwagie, J., Janssens, W., Piot, P., and van der Groen, G. (1995) Design and evaluation of new, highly sensitive and specific primers for polymerase chain reaction detection of HIV-1 infected primary lymphocytes. Mol. Cell. Probes 9, 373. 

Rychlik, W. 1993. Selection of primers for polymerase chain reaction. In Methods in molecular biology, Vol. 15 PCR protocols Current methods and applications, ed. B. A. White, 31 40. Totowa, NJ Humana. 

Ye, J., Coulouris, G., Zaretskaya, I., Cutcutache, L, Rozen, S., Madden, T. (2012). Primer-BLAST a tool to design target-specific primers for polymerase chain reaction. BMC Bioinformatics, 13,134. 

Differential display is a method for identifying differentially expressed genes, using anchored oligo-dT, random oligonucleotide primers and polymerase chain reaction on reverse-transcribed RNA from different cell populations. The amplified complementary DNAs are displayed and comparisons are drawn between the different cell populations. 

Figure 9.4. Request form for primer selection. The nucleotide sequence of a target DNA for polymerase chain reaction can be submitted for primer selection at Primer3 server.

Zoh GJ, Melchers WJ, Kopecka H, Jambroes G, van der Poel HJ, Galaraa JM. General primer-mediated polymerase chain reaction for detection of enteroviruses application for diagnostic routine and persistent infections. J Clm Microbiol 1999 30 60-5. 

To this end, cDNA libraries, prepared from cells or tissues known to be rich in certain receptors, were screened by low stringency hybridization, or were used for polymerase chain reaction (PCR) amplification of candidate genes using degenerate primers. Proof of function was obtained after the... 

Fig. 6. Polymerase chain reaction (PCR) mediated site-directed mutagenesis. The 5 and 3 ends of the nucleotide strands are indicated. The four arrows surrounding the DNA template represent oligonucleotide primers 1—4. See text for discussion.

While many diseases have long been known to result from alterations in an individual s DNA, tools for the detection of genetic mutations have only recently become widely available. These techniques rely upon the catalytic efficiency and specificity of enzyme catalysts. For example, the polymerase chain reaction (PCR) relies upon the ability of enzymes to serve as catalytic amplifiers to analyze the DNA present in biologic and forensic samples. In the PCR technique, a thermostable DNA polymerase, directed by appropriate oligonucleotide primers, produces thousands of copies of a sample of DNA that was present initially at levels too low for direct detection. 

Fig. 19.1 Differential displays comparing RNAs from saline (S)-, imipramine (I)- or fluoxetine (F)-treated rats. Total RNA was extracted from hypothalami of animals treated with the different drugs for two months. Autoradiograms of amplified -[35S]-dATP-labeled PCR (polymerase chain reaction) products after electrophoresis in 6% polyacrylamide gels are shown for two different primer combinations that identified one upregulated (arrowhead) and one downregulated (arrow) fragment in the groups treated with antidepressants (from [4] with permission).

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