Accurate and highly sensitive screening of DNA-binding molecules with primer-dimer templates using a real-time PCR analyzer.

نویسندگان

  • Akira Inoue
  • Tsunehiro Kumeda
  • Hiroshi Nakamura
چکیده

We have developed a strategy to detect the DNA-binding affinities of chemical compounds with a real-time PCR analyzer. In this system, the DNA-binding affinities of chemical compounds are efficiency detected by PCR with high sensitivity. To obtain accurate information about the DNA-binding affinities of chemical compounds, primer-dimers prepared in advance are used as templates. The denature temperature is settled to around the melting temperature of the templates.

برای دانلود رایگان متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Development of a Sensitive Quantitative Competitive PCR Assay for Detection of Human Cytomegalovirus DNA

Accurate and rapid diagnosis of human cytomegalovirus (HCMV) disease in immunocompromised patients has remained as a challenge. Quantitative competitive PCR (QC-PCR) methods for detection of HCMV in these individuals have improved the positive and negative predictive values of PCR for diagnosis of HCMV disease. In this study we used QC-PCR assay, using a co-amplified DNA standard, to quantitate...

متن کامل

Application of Rapid and Sensitive Real Time PCR Technique in Detection of DNA Impurities in Recombinant Interferon

 Background & Objective: Interferon belongs to a family of cytokines, which has the most important role in the innate immune response to virus infections. While producing recombinant interferon in biological host, some pieces of host nucleic acids remain in product. Because of limitations in previous techniques for detection of these impurities, the objective of this study is to use rapid ...

متن کامل

Development of SYBR Green I Based Real-Time RT-PCR Assay for Specific Detection of Watermelon silver mottle Virus

Background: Watermelon silver mottle virus (WSMoV), which belongs to the genus Tospovirus, causes significant loss in Cucurbitaceae plants. Objectives: Development of a highly sensitive and reliable detection method for WSMoV. Materials and Methods: Recombinant plasmids for targeting the sequence of nucleocapsid protein gene of WSMoV were constructed. SYBR Green I real-time PCR was established...

متن کامل

Analysis of Herpes Simplex Virus in Suspected Encephalitis, Keratitis and Dermal Infections Using Real- Time PCR

Background & Objectives: Herpes viruses can cause diseases in the clinical range. The virus can cause infection in various body parts, especially eyes and nervous system. The aim of this study was at evaluating  the Real-Time TaqMan probe PCR in diagnosing and monitoring of the patients with suspected HSV infections.Materials & Methods: More than a thousand patients with suspected HSV infection...

متن کامل

Elimination of primer-dimer artifacts and genomic coamplification using a two-step SYBR green I real-time RT-PCR.

Gene expression analysis plays an increasingly important role in many fields of biological research. The recently developed real-time PCR quantification method has many advantages over the conventional quantifications in terms of accuracy, sensitivity, dynamic range, high-throughput capacity, and absence of post-PCR manipulations (1, 2). Sequence-specific fluorescence-labeled probes (e.g., TaqM...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

عنوان ژورنال:
  • Analytical sciences : the international journal of the Japan Society for Analytical Chemistry

دوره 27 5  شماره 

صفحات  -

تاریخ انتشار 2011