qScript One-Step RT-qPCR Kit

Features & Benefits

  • 1-step convenience
  • high efficiency and sensitivity
  • available with reference dye pre-mixed

qScript One-Step RT-qPCR Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention or treatment of a disease.

Description

The qScript One-Step qRT-PCR Kit is a convenient and highly sensitive solution for reverse transcription quantitative PCR (RT-qPCR) of RNA templates using hybridization probe detection chemistries such as TaqMan(r) 5'-hydrolysis probes.  cDNA Synthesis and PCR amplification are carried out in the same tube without opening between procedures.  The system has been optimized to deliver maximum RT-PCR efficiency, sensitivity, and specificity, enabling unbiased co-amplification of low copy transcripts in the presence of higher copy reference genes.  The proprietary reaction buffer has been specifically formulated to maximize activities of both reverse transcriptase and Taq DNA polymerase while minimizing the potential for primer-dimer and other non-specific PCR artifacts.

Details

  • Contents

    qScript One-Step Reverse Transcriptase (50x)

    One-Step Master Mix 2x (optionally containing ROX or Low ROX)

    Nuclease-free water

  • Storage & Handling
    Store components in a constant temperature freezer at -25°C to -15°C protected from light upon receipt. Repeated freezing and thawing of the reaction mix is not recommended. For lot specific expiry date, refer to package label, Certificate of Analysis or Product Specification Form.
  • Instrument Capability
    ROX
    • Applied Biosystems 5700
    • Applied Biosystems 7000
    • Applied Biosystems 7300
    • Applied Biosystems 7700
    • Applied Biosystems 7900
    • Applied Biosystems 7900HT
    • Applied Biosystems 7900 HT Fast
    • Applied Biosystems StepOne™
    • Applied Biosystems StepOnePlus™
    Low ROX
    • Applied Biosystems 7500
    • Applied Biosystems 7500 Fast
    • Stratagene Mx3000P®
    • Stratagene Mx3005P™
    • Stratagene Mx4000™
    • Applied Biosystems ViiA 7
    • Applied Biosystems QuantStudio™
    • Agilent AriaMx
    • Douglas Scientific IntelliQube®
    No ROX
    • Quantabio Q
    • BioRad CFX
    • Roche LightCycler 480
    • QIAGEN Rotor-Gene Q
    • Other
    Bio-Rad iCycler iQ systems
    • BioRad iCycler iQ™
    • BioRad MyiQ™
    • BioRad iQ™5
  • Related Resources
    Product Manuals
    Technical Notes
    FAQs
    What is the amount of RNA used per TaqMan® or SYBR qPCR Assay?
    Suggested input quantities of template are: 1 pg to 1 μg total RNA; 10 fg to 100 ng poly A(+) RNA; 10 to 1x108 copies viral RNA. For more information, please consult the qScrip One-Step qRT-PCR Kits protocols.
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    Publications
    Soluble Fn14 Is Detected and Elevated in Mouse and Human Kidney Disease
    M. Nusrat Sharif, PLOS ONE - 2016
    Abstract
    The cytokine TWEAK and its cognate receptor Fn14 are members of the TNF/TNFR superfamily and are upregulated in tissue injury to mediate local tissue responses including inflammation and tissue remodeling. We found that in various models of kidney disease, Fn14 expression (mRNA and protein) is upregulated in the kidney. These models include: lupus nephritis mouse models (Nephrotoxic serum Transfer Nephritis and MRL.Fas lpr/lpr ), acute kidney injury models (Ischemia reperfusion injury and Folic acid injury), and a ZSF-1 diabetic nephropathy rat model. Fn14 expression levels correlate with disease severity as measured by disease histology. We have also shown for the first time the detection of soluble Fn14 (sFn14) in the urine and serum of mice. Importantly, we found the sFn14 levels are markedly increased in the diseased mice and are correlated with disease biomarkers including proteinuria and MCP-1. We have also detected sFn14 in human plasma and urine. Moreover, sFn14 levels, in urine are significantly increased in DN patients and correlated with proteinuria and MCP-1 levels. Thus our data not only confirm the up-regulation of Fn14/TWEAK pathway in kidney diseases, but also suggest a novel mechanism for its regulation by the generation of sFn14. The correlation of sFn14 levels and disease severity suggest that sFn14 may serve as a potential biomarker for both acute and chronic kidney diseases.
    Overwintering of Rabies Virus in Silver Haired Bats ( Lasionycteris noctivagans )
    April Davis, PLOS ONE - 2016
    Abstract
    Silver-haired bats, ( Lasionycteris noctivagans ) are semi-colonial, migratory tree bats that have infrequent contact with humans. Despite the species rarity, the L . noctivagans rabies variant is the most commonly reported rabies virus variant (RABV) in domestically acquired human rabies cases in the US. Unlike big brown bats ( Eptesicus fuscus ) and little brown bats ( Myotis lucifugus ), L . noctivagans are not considered true hibernators. It is unknown if RABV can overwinter in hibernating L . noctivagans or is only maintained in members of this taxa that migrate to warmer climates. To better understand RABV overwintering in this species, L . noctivagans were inoculated intramuscularly with either a homologous RABV ( L . noctivagans Virus 1) or one of two heterologous RABV ( Eptesicus fuscus Virus 2 and Myotis lucifugus Virus 1). Five days following inoculation, L . noctivagans were placed in a hibernation chamber for 6 weeks. Our results demonstrate that rabies virus can overwinter in L . noctivagans yet the incubation period was extended 6 weeks when compared to bats maintained at ambient temperatures. Additionally, we found that the longer the incubation period, the greater the viral dissemination to the salivary glands. Similar to our previous studies, L . noctivagans were most susceptible to a homologous variant. In summary, we found that RABV incubation is extended following a subcutaneous exposure or maintenance in hibernation and longer incubation times increase dissemination and potential for transmission.
    DENGUE VIRUS (DV) POLYPEPTIDE SEQUENCES, T CELL EPITOPES AND METHODS AND USES THEREOF - LA JOLLA INSTITUTE FOR ALLERGY AND IMMUNOLOGY
    Sujan Shresta, United States Patent - 2016
    Abstract
    Dengue virus (DV) peptides, including T cell epitopes, structural and non-structural (NS) polypeptide sequences, subsequences and modifications thereof, nucleotide sequences encoding such peptides, and compositions including such peptides and encoding nucleotide sequences, and cells expressing such peptides, are provided. Such DV peptides, nucleotide sequences and compositions, can be used to elicit, stimulate, induce, promote, increase, enhance or activate an anti-DV CD8+ T cell response or an anti-DV CD4+ T cell response. Such peptides, nucleotide sequences and compositions can also be used for and in methods of vaccination/immunization of a subject against Dengue virus (DV) (e.g., to provide protection against DV infection and/or pathology), and for treatment of a subject in need thereof, for example, treatment of the subject for a Dengue virus (DV) infection or pathology.
    Control of Citrus Huanglongbing (HLB) via Trunk Injection of Plant Activators and Antibiotics
    Jiahuai Hu, Phytopathology - 2017
    Abstract
    Citrus Huanglongbing (HLB) or greening is a devastating disease of citrus worldwide and no effective control measure is currently available. Plant activators represent environment friendly compounds capable of inducing resistance against many plant pathogens. Earlier studies showed that foliar spray of plant defense inducers could slow down HLB disease progress. In this study, eight plant activators and three antibiotics were evaluated in 3 field trials for their effect to control HLB by trunk injection of young and mature sweet orange trees. Results showed that 4 trunk injections of several activators including salicylic acid, oxalic acid, acibenzolar-S-methyl and potassium phosphate provided significant control of HLB by suppressing Las titer and disease progress. Trunk injection of penicillin, streptomycin and oxytetracycline hydrochloride resulted in excellent control of HLB. In general, antibiotics were more effective in reduction of Las titer and HLB symptom expressions than plant activators. These treatments also resulted in increased yield and better fruit quality. Injection of both salicylic acid and acibenzolar-S-methyl led to significant induction of PR-1 and PR-2 genes. Meanwhile, injection of either potassium phosphate or oxalic acid resulted in significant induction of PR-2 or PR-15 gene expression, respectively. These results suggested that HLB diseased trees remained inducible for systemic acquired resistance (SAR) under field conditions. In summary, this study presents information regarding controlling HLB via trunk injection of plant defense activators and antibiotics, which helps citrus growers in decision-making regarding developing an effective HLB management program.
    CD4+ T cells promote humoral immunity and viral control during Zika virus infection
    Annie Elong Ngono, PLOS ONE - 2019
    Abstract
    Several Zika virus (ZIKV) vaccines designed to elicit protective antibody (Ab) responses are currently under rapid development, but the underlying mechanisms that control the magnitude and quality of the Ab response remain unclear. Here, we investigated the CD4+ T cell response to primary intravenous and intravaginal infection with ZIKV. Using the LysMCre+Ifnar1fl/fl (myeloid type I IFN receptor-deficient) C57BL/6 mouse models, we identified six I-Ab-restricted ZIKV epitopes that stimulated CD4+ T cells with a predominantly cytotoxic Th1 phenotype in mice primed with ZIKV. Intravenous and intravaginal infection with ZIKV effectively induced follicular helper and regulatory CD4+ T cells. Treatment of mice with a CD4+ T cell-depleting Ab reduced the plasma cell, germinal center B cell, and IgG responses to ZIKV without affecting the CD8+ T cell response. CD4+ T cells were required to protect mice from a lethal dose of ZIKV after infection intravaginally, but not intravenously. However, adoptive transfer and peptide immunization experiments showed a role for memory CD4+ T cells in ZIKV clearance in mice challenged intravenously. These results demonstrate that CD4+ T cells are required mainly for the generation of a ZIKV-specific humoral response but not for an efficient CD8+ T cell response. Thus, CD4+ T cells could be important mediators of protection against ZIKV, depending on the infection or vaccination context.
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    CofA (PSF)