Thermostable DNA polymerase gene from Thermus aquaticus was cloned into constructed Taq from Thermus a Qaticus (pTTQ) plasmid using EcoRI and SalI sites with subsequent transformation in Escherichia coli strain (TOP10). The use of Isopropyl-β-Dthiogalactopyranosid (IPTG) as inducer of interested gene expression under control of the lac promoter was investigated. The optimization of enzyme induc...

For the detection of RNA transcripts by RT-PCR, prior removal of genomic DNA must be performed. To remove genomic DNA, RNA is often prepared by DNase I digestion following phenol extraction. Recently, one-tube or onebuffer systems of RT-PCR were developed to prevent loss of RNA and to reduce the risk of contamination (2,6). In these methods, DNase I is added before RT. Taq DNA polymerase prepar...

The full potential of diagnostic PCR is limited, in part, by the presence of inhibitors in complex biological samples that reduce the amplification efficiency. Therefore, different pre-PCR treatments are being used to reduce the effects of PCR inhibitors. The aim of the present study was to investigate the effects of 16 amplification facilitators to enhance DNA amplification in the presence of ...

Plasma concentrations of HDL (high-density lipoprotein) cholesterol are low in the Saudi Arabian population. A B polymorphism at the CETP (cholesteryl ester protein transfer) locus that is detectable with the restriction enzyme Taq I is a genetic determinant of the plasma HDL cholesterol concentration. We assessed the relationship between the Taq I B CETP polymorphism and lipid and apolipoprote...

For certain applications of the polymerase chain reaction (PCR), it may be necessary to consider the accuracy of replication. The breakthrough that made PCR user friendly was the commercialization of Thermus aquaticus (Taq) DNA polymerase, an enzyme that would survive the high temperatures needed for DNA denaturation. The development of enzymes with an inherent 3' to 5' exonuclease proofreading...

Important in understanding the regulation of gene transcription is the elucidation of specific protein-DNA interactions that occur in vivo. One strategy for such in vivo footprinting involves the treatment of whole cells with dimethyl sulphate (DMS), which leads to methylation of guanine residues in DNA at the N7 position (1, 2). This N7 atom lies in the major groove and its susceptibility to m...

AIM To investigate the dose-response effect of humic acid (HA) on the quantitative real time polymerase chain reaction (QRT-PCR) inhibition and the efficiency of Taq polymerase increment in preventing inhibition by HA in DNA extracted from ancient bones. METHODS DNA was isolated from bone samples and DNA quantification was conducted with the real-time 5' exonuclease detection assay (TaqMan), ...

Different amplification protocols were evaluated for use with primer-extension preamplification (PEP). We hypothesized that a protocol known to improve amplification of long DNA fragments would improve efficacy of PEP. Eight DNA samples were preamplified by PCR using different protocols. Treatments consisted of the use of Taq DNA polymerase (T), Taq plus a second polymerase obtained from Pyroco...

The concept of acceptance has recently received growing attention within tinnitus research due to the fact that tinnitus acceptance is one of the major targets of psychotherapeutic treatments. Accordingly, acceptance-based treatments will most likely be increasingly offered to tinnitus patients and assessments of acceptance-related behaviours will thus be needed. The current study investigated ...