The AA genotype of the regulatory BCL2 promoter polymorphism (938C>A) is associated with a favorable outcome in lymph node negative invasive breast cancer patients. Characterization of the GNAQ promoter and association of increased Gq expression with cardiac hypertrophy in humans. A novel promoter polymorphism in the human gene GNAS affects binding of transcription factor upstream stimulatory factor 1, Galphas protein expression and body weight regulation. Successful amplification of extremely GC-rich promoter regions using a novel 'slowdown PCR' technique. Regulation of gene expression by GC-rich DNA cis-elements. Denaturants or cosolvents improve the specificity of PCR amplification of a G + C-rich DNA using genetically engineered DNA polymerases. Localised sequence regions possessing high melting temperatures prevent the amplification of a DNA mimic in competitive PCR. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction. (1991) Excessive cycling converts PCR products to random-length higher molecular weight fragments. (1991) Recent advances in the polymerase chain reaction. (1991) Maximizing sensitivity and specificity of PCR by preamplification heating. (1994) New algorithm for determining primer efficiency in PCR and sequencing. (1989) A computer program for choosing optimal oligonucleotides for filter hybridization, sequencing and in vitro amplification of DNA. (1993) PCR with degenerate primers containing deoxyinosine fails with Pfu DNA polymerase. (1991) Structure and functional properties of human general transcription factor IIE. E., Flores, O., Adomon, A., Reinberg, D., and Tjian, R. (1995) Optimization and troubleshooting in PCR. (1991) Enhanced evolutionary PCR using oligonucleotides with inosine at the 3′-terminus. (1990) PCR amplification of an Escherichia coli gene using mixed primers containing deoxyinosine at ambiguous positions in degenerate amino acid codons. (1988) Highly degenerate inosine-containing primers specifically amplify rare cDNA using the polymerase chain reaction. Knoth, K., Roberds, S., Poteet, C., and Tamkun, M. (1994) Using mismatched primer-template pairs in TD PCR. (1996) High and low annealing temperatures increase both specificity and yield in TD and SD PCR. (1991) ‘Touchdown’ PCR to circumvent spurious priming during gene amplification. This process is experimental and the keywords may be updated as the learning algorithm improves.ĭon, R. These keywords were added by machine and not by the authors. Unfortunately, even with the most sophisticated algorithms (i.e., OLIGO) it is often difficult to predict the amplification optima a priori leaving no other choice but to employ empirical determination. With regard to the latter, the value selected for the annealing temperatures is most critical. Variables include concentrations of Mg 2+, H +, dNTPs, primers, and template, as well as cycling parameters. Reactions that are too stringent yield negligible product and reactions that are not stringent enough yield artifactual amplicons. Small variations in any of the many variables in a given reaction can have a pronounced effect on the resultant amplicon profile. Polymerase chain reaction (PCR) optimization and troubleshooting can consume considerable energy and resources because of the finicky and often unpredictable nature of the reactions.
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