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PerfeCTa qPCR SuperMix

Features & Benefits
Product
Kit Size
Order Info
Product
Kit Size
Order Info
PerfeCTa qPCR SuperMix
Request Sample
Kit Size:
Order Info:
500 x 50 μL rxns (10 x 1.25 mL)
2000 x 50 μL rxns (1 x 50 mL)
Product
Kit Size
Order Info
PerfeCTa qPCR SuperMix ROX
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Kit Size:
Order Info:
500 x 50 μL rxns (10 x 1.25 mL)
2000 x 50 μL rxns (1 x 50 mL)
Product
Kit Size
Order Info
PerfeCTa qPCR SuperMix Low-ROX
Request Sample
Kit Size:
Order Info:
500 x 50 μL rxns (10 x 1.25 mL)
2000 x 50 μL rxns (1 x 50 mL)

Description

PerfeCTa qPCR SuperMix is a 2X concentrated, ready-to-use reaction cocktail that contains all components, except primers, probe(s), and template for real-time quantitative PCR systems. The proprietary buffer and stabilizers have been specifically optimized to deliver maximum PCR efficiency, sensitivity, and robust fluorescent signal with TaqMan® or TaqMan MGB probe chemistry. The enhanced specificity of this supermix suppresses cross-reactivity between homologous sequences, improving detection and discrimination in SNP applications. A key component of this supermix is AccuStart Taq DNA polymerase, which contains monoclonal antibodies that bind to the polymerase and keep it inactive prior to the initial PCR denaturation step. Upon heat activation (2 minutes at 95ºC), the antibodies denature irreversibly, releasing fully active, unmodified Taq DNA polymerase. This enables specific and efficient primer extension with the convenience of room temperature reaction assembly.

Details

Details

Contents

PerfeCTa qPCR SuperMix (2X): 2X reaction buffer containing optimized concentrations of MgCl2, dNTPs (dATP, dCTP, dGTP, dTTP), AccuStart Taq DNA Polymerase, and stabilizers.

Instrument Capability

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

Details

Contents

PerfeCTa qPCR SuperMix (2X): 2X reaction buffer containing optimized concentrations of MgCl2, dNTPs (dATP, dCTP, dGTP, dTTP), AccuStart Taq DNA Polymerase, and stabilizers.

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

Resources

Product Manuals

CofA (PSFs)

Click here to see all CofA (PSFs)

SDSs

Publications

Role of Diospyros peregrina fruit preparation in suppressing regulatory T (Treg) cells in the tumor microenvironment of breast and lung cancer
Rishabh Rai Kaushik - 2022
Abstract
Background The most frequent cancers in the world are breast cancer and lung cancer. Regulatory T cells (Treg) are the subtype of T cells that promotes equilibrium between immunological surveillance and immune tolerance, and they are commonly found in high amounts within tumours. The presence of Treg cells in the tumour microenvironment results in immune suppression, due to which immune cells are not able to kill the tumour cells. Diospyros peregrina (Gaertn.) Gürke is an ethnomedicinal plant with hypoglycaemic, diuretic, and anti-cancer effects. This genus, Diospyros, is native to India but distributed throughout the tropics and is characterized by its ability to produce triterpenes of the lupine series and various other bioactive compounds. Diospyros peregrina fruit preparation (DFP) substantially increases tumour protective immunity by suppressing Treg cells. Purpose This project aims to suppress the regulatory T cells in the tumour microenvironment by giving Diospyros peregrina fruit preparation (DFP) treatment to Treg cells, which will help in controlling and combating breast cancer and lung cancer. Method For this project's work, we isolated peripheral blood mononuclear cells from the blood, and the isolated peripheral blood mononuclear cells were used to isolate CD4+CD25+ Treg cells. Then, Diospyros peregrina fruit preparation (DFP) was prepared, which was used to treat CD4+CD25+ Treg cells. Lactate dehydrogenase (LDH) assays, enzyme-linked immunosorbent assays (ELISA), and RT-qPCR were used to determine the effect of DFP treatment on Treg cells. Result Diospyros peregrina fruit preparation (DFP) treatment suppresses the expression of CD4+CD25+ Foxp3+Treg cells within tumours. DFP-treated CD4+CD25+Foxp3+ Treg cells increase the cytotoxicity within the tumours compared to non-treated Treg cells. DFP treatment help in reducing the release of (T helper type 2) TH2 cell cytokines (Interleukin-10) IL-10 as well as (Interleukin-4) IL-4 and escalating the release of (T helper type 1) TH1 cell cytokines (Interleukin-12) IL-12 and (Interferon-γ) IFN-γ, it also increases the transcriptional activity of T-box transcription factor TBX21 (TBX21) and reduces the expression of the transcription factor FOXP3 and GATA3. Conclusion Diospyros peregrina fruit preparation (DFP) treatment augments the expressions of TH1-specific cytokines and transcription factors, and DFP treatment diminishes the expressions of both TH2 and Treg-specific cytokines and transcription factors.
p53 regulates skeletal muscle mitophagy and mitochondrial quality control following denervation-induced muscle disuse
Jonathan M. Memme - 2021
Abstract
Persistent inactivity promotes skeletal muscle atrophy, marked by mitochondrial aberrations that affect strength, mobility, and metabolic health leading to the advancement of disease. Mitochondrial quality control (MQC) pathways include biogenesis (synthesis), mitophagy/lysosomal turnover, and the mitochondrial unfolded protein response (UPRmt) which serve to maintain an optimal organelle network. Tumor suppressor p53 has been implicated in regulating muscle mitochondria in response to cellular stress; however, its role in the context of muscle disuse has yet to be explored, and whether p53 is necessary for MQC remains unclear. To address this, we subjected p53 muscle-specific knockout (mKO) and wild-type (WT) mice to unilateral denervation. Transcriptomic and pathway analyses revealed dysregulation of pathways pertaining to mitochondrial function, and especially turnover, in mKO muscle following denervation. Protein and mRNA data of the MQC pathways indicated activation of the UPRmt and mitophagy-lysosome systems along with reductions in mitochondrial biogenesis and content in WT and mKO tissue following chronic denervation. However, p53 ablation also attenuated the expression of autophagy/mitophagy machinery, reduced autophagic flux, and enhanced lysosomal dysfunction. While similar reductions in mitochondrial biogenesis and content were observed between genotypes, MQC dysregulation exacerbated mitochondrial dysfunction in mKO fibers, evidenced by elevated reactive oxygen species (ROS). Moreover, acute experiments indicate that p53 mediates the expression of transcriptional regulators of MQC pathways as early as 1 day following denervation. Together, our data illustrate exacerbated mitochondrial dysregulation with denervation stress in p53 mKO tissue, thus indicating that p53 contributes to organellar maintenance via regulation of MQC pathways during muscle atrophy.
Differential expression of Oct3/4 in human breast cancer and normal tissues
Feng-Qi Zhao - 2018
Abstract
Oct3/4, a transcription factor specifically expressed in mammalian totipotent embryonic stem and germ cells, has a critical role in the regulation and maintenance of pluripotency and self-renewal. However, reactivation of Oct3/4 expression is observed in several human breast cancer cell lines, but not in non‑malignant cells. To examine Oct3/4 expression in human primary breast carcinomas and normal breast tissues, we obtained breast tumor tissues from 28 patients and normal breast tissues from 9 women. According to quantitative polymerase chain reaction, all of the tumor tissues, irrespective of tumor type or clinicopathological status, expressed Oct3/4 mRNA at 10- to 100- fold higher levels than that in the normal breast tissues. Expression of the Oct3/4 protein in tumors was confirmed by western blot analysis and immunofluorescent staining. Additionally, rapid amplification of cDNA ends and DNA sequencing revealed expression of multiple Oct4 gene transcripts from chromosome 6 (POU5F1) in normal breast tissues and the non‑malignant breast epithelial cell line MCF‑10A; by contrast, the breast tumors and malignant breast cancer cell line MCF‑7 predominantly expressed transcripts of an Oct4-like gene (POU5F1B) from chromosome 8, which was termed Oct3 in the current study. The deduced amino acid sequences of full-length Oct3 and Oct4 are 96% identical. The findings of the current study indicated that Oct3, rather than Oct4, may serve as a novel clinical marker and a potential target for gene-specific therapy of breast cancer
Establishment of neomycin phosphotransferase II
Asuka Itaya - 2018
Abstract
Soybean transformation is limited by the lack of multiple efficient selectable marker systems. Biolistic transformation of somatic proliferative embryogenic cultures, one of the commonly used soybean transformation methods, relies largely on hygromycin phosphotransferase II (hptII) selection. The purpose of the present study was to establish another efficient selectable marker system to facilitate multiple gene transformations of soybean. We tested neomycin phosphotransferase II (nptII) that has been used successfully in cotyledonary node transformation, but with limited success in transformation of embryogenic cultures. Transgenic events were obtained using nptII with improved G418 selection without generating escapes. G418 selection required longer recovery and selection periods, and resulted in a lower efficiency of initial transformants compared to hygromycin selection. Six independent fertile transgenic plants were recovered using nptII and G418, a frequency similar to that obtained with hygromycin selection. Soybean embryogenic cultures co-transformed with the hptII and nptII markers showed resistance to both hygromycin B and G418, while regeneration and plant fertility were not adversely affected. The nptII will be useful as a second selectable marker for multiple gene transformations in basic and applied soybean research.

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  • 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®
  • 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™
  • Quantabio Q
  • BioRad CFX
  • Roche LightCycler 480
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