When dCTP is replaced by methyl5-dCTP in the polymerase chain reaction some templates cannot be efficiently amplified by Taq polymerase or Vent polymerase using standard cycling parameters. However, this phenomenon can be overcome by increasing the temperature of the denaturation steps to 100 degrees C, or by adding dITP to destabilize the m5dC:dG base pairs. Once the block to amplification of ...

The polymerase chain reaction (PCR) has revolutionized the molecular biology industry, with profound implications for medical, forensic and research sciences. Prerequisites for this technology to work include the presence of deoxynucleotriphosphates (dNTPs), a relevant buffering system, magnesium chloride, primers and template DNA and finally, the DNA polymerase. Taq polymerase, a thermostable ...

DNase I pretreatment of 16S rRNA gene PCR reagents was tested. The DNase I requirement for the elimination of false-positive results varied between 0.1 and 70 IU per master mix depending on the applied Taq polymerase. PCR sensitivity was mostly maintained when 0.1 IU of DNase I was used.

The carryover of previously amplified sequences (amplicons) into new PCR reactions is a serious problem. Recently Furrer et al. reported a 'pre-PCR' sterilization technique using DNase I or restriction enzymes for contamination control (1). Unfortunately, these methods require the reaction tube to be re-opened to add target DNA and Taq polymerase following the enzymatic treatment (providing an ...

Objectives . This study investigated the substrate properties of modified derivatives triphosphates purine and pyrimidine deoxynucleosides (5-propynyl-2’-deoxyuridine-5’-triphosphate, 5-propynyl2’-deoxycytidine-5’-triphosphate, 5-methyl-2’-deoxycytidine-5’-triphosphate, N6-methyl-2’-deoxyadenosine-5’-triphosphate) during their simultaneous incorporation in enzymatic reactions (polymerase chain ...

Fonsecin B has been identified as stabilizing ligand of c-myc G-quadruplex DNA using high-throughput virtual screening of a natural product database, and inhibited Taq polymerase-mediated DNA extension in vitro through stabilization of the G-quadruplex secondary structure.

A PCR procedure has been developed for routine analysis of viable Salmonella spp. in feed samples. The objective was to develop a simple PCR-compatible enrichment procedure to enable DNA amplification without any sample pretreatment such as DNA extraction or cell lysis. PCR inhibition by 14 different feed samples and natural background flora was circumvented by the use of the DNA polymerase Tth...