164 research outputs found

    A single amino acid change to Taq DNA polymerase enables faster PCR, reverse transcription and strand-displacement

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    A change of an aspartic acid to asparagine of Taq

    Investigation of 6TSWCNT by Cs-Corrected Transmission Electron Microscopy

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    Extended abstract of a paper presented at Microscopy and Microanalysis 2009 in Richmond, Virginia, USA, July 26 - July 30, 200

    Mutants of Taq DNA polymerase resistant to PCR inhibitors allow DNA amplification from whole blood and crude soil samples

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    Potent PCR inhibitors in blood and soil samples can cause false negative results from PCR-based clinical and forensic tests. We show that the effect of these inhibitors is primarily upon Taq DNA polymerase, since mutational alteration of the polymerase can overcome the inhibition to the extent that no DNA purification is now required. An N-terminal deletion (Klentaq1) is some 10ā€“100-fold inhibition resistant to whole blood compared to full-length, wild-type (w.t.) Taq, which is strongly inhibited by 0.1ā€“1% blood. Further mutations at codon 708, both in Klentaq 1 and Taq, confer enhanced resistance to various inhibitors of PCR reactions, including whole blood, plasma, hemoglobin, lactoferrin, serum IgG, soil extracts and humic acid, as well as high concentrations of intercalating dyes. Blood PCR inhibitors can predominantly reduce the DNA extension speed of the w.t. Taq polymerase as compared to the mutant enzymes. Single-copy human genomic targets are readily amplified from whole blood or crude soil extract, without pretreatment to purify the template DNA, and the allowed increase in dye concentration overcomes fluorescence background and quenching in real-time PCR of blood

    Androgen deprivation decreases prostate specific antigen in the absence of tumor: implications for interpretation of PSA results

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    Background: Prostate-specific antigen (PSA) is used as an outcome measure for relapsed disease in prostate cancer. Nonetheless, there are considerable concerns about its indiscriminate use as a surrogate endpoint for cell growth or survival. We hypothesized that treatment with a luteinizing hormone releasing hormone (LHRH) analog would decrease PSA levels even in the absence of malignant disease. Methods: We determined testosterone and PSA levels in 30 healthy volunteers after a single intramuscular injection of a LHRH depot formulation. Testosterone and PSA levels were quantified by radioimmunoassay and electrochemi-luminescence immunoassay, respectively. Results: After an initial flare-up during the first 3 days testosterone decreased reaching castration levels in 18 of the 30 young men (60%). After the nadir on day 28, testosterone levels increased to normal again. Changes in PSA paralleled those of testosterone. Castration reduced PSA levels by 29% (95% CI 19%-39%) compared to baseline (p<0.0001). Conclusions: LHRH superagonists decrease PSA levels by testosterone deprivation. Conferring these findings to tumor patients, decreases in PSA after treatment with LHRH analogs might not only reflect disease regression but also a direct testosterone mediated effect on PSA. Thus, PSA levels should be cautiously interpreted when patients receive hormonal therap

    Investigation of 6T@SWNTs by Cs corrected TEM

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    Origin of the Superconductivity in the Y-Sr-Ru-O and Y-Sr-Cu-O Systems

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    We report on the structural, magnetic, and Raman-scattering studies of double perovskite structure Sr2Y(Ru1-xCux)O6-d systems made by systematic synthesis processes with various numbers of doping concentrations and sintering temperatures. We observed different behaviors resulting from the different thermal treatments. In particular, superconductivity in Cu-doped Sr2YRuO6 has been observed only for partially melted ceramic materials. We show that superconductivity is associated with the 1:2:3 phase (YSr2Cu3Ot), similar to that of Y-Sr-Cu-O samples sintered at high temperature

    The evaluation of radiation damage parameter for CVD diamond

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    There are a few different phenomenological approaches that aim to track the dependence of signal height in irradiated solid state detectors on the fluence of damaging particles. However, none of them are capable to provide a unique radiation hardness parameter that would reflect solely the material capability to withstand high radiation environment. To extract such a parameter for chemical vapor deposited (CVD) diamond, two different diamond detectors were irradiated with proton beams in MeV energy range and subjected afterwards to ion beam induced charge (IBIC) analysis. The change in charge collection efficiency (CCE) due to defects produced was investigated in context of a theoretical model that was developed on the basis of the adjoint method for linearization of the continuity equations of electrons and holes. Detailed modeling of measured data resulted with the first known value of the kr product for diamond, where k represents the number of charge carriersā€™ traps created per one simulated primary lattice vacancy and r represents the charge carriersā€™ capture cross section. As discussed in the text, this product could be considered as a true radiation damage parameter
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