Quantification of Murine IFN-γ mRNA and Protein Expression: Impact of Real-Time Kinetic RT-PCR Using SYBR Green I Dye
نویسندگان
چکیده
منابع مشابه
Accurate and statistically verified quantification of relative mRNA abundances using SYBR Green I and real-time RT-PCR.
Among the many methods currently available for quantifying mRNA transcript abundance, reverse transcription-polymerase chain reaction (RT-PCR) has proved to be the most sensitive. Recently, several protocols for real-time relative RT-PCR using the reporter dye SYBR Green I have appeared in the literature. In these methods, sample and control mRNA abundance is quantified relative to an internal ...
متن کاملDevelopment 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 کاملAbsolute quantification of murine interleukine-4, interleukine- 10 and interferon-γ gene transcripts using Real Time PCR
The study of cytokines gene expression is quite important in various conditions of health and disease for the evaluation of clinical responses to new vaccination approaches. An absolute quantification is based on a calibration curve and production of standard controls to achieve more reliable results than in relative system. In this study we attempted to construct standard controls to evaluate ...
متن کاملA Broadly Reactive One-Step SYBR Green I Real-Time RT-PCR Assay for Rapid Detection of Murine Norovirus
A one-step SYBR Green I real-time RT-PCR assay was developed for the detection and quantification of a broad range of murine noroviruses (MNVs). The primer design was based on the multiple sequence alignments of 101 sequences of the open reading frame (ORF)1-ORF2 junction of MNV. The broad reactivity and quantitative capacity of the assay were validated using 7 MNV plasmids. The assay was compl...
متن کاملElimination 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 کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
ژورنال
عنوان ژورنال: Scandinavian Journal of Immunology
سال: 2001
ISSN: 0300-9475
DOI: 10.1046/j.1365-3083.2001.00928.x