PerfeCTa SYBR<sup>®</sup> Green SuperMix

PerfeCTa SYBR^® Green SuperMix

Sensitive and precise DNA amplification with DNA-intercalating dye based detection chemistry

Features & Benefits

2x concentrated reagents minimize pipetting steps, simplify reaction assembly and improve accuracy
Superior assay sensitivity and specificity with ultrapure AccuStart enzyme technology – maximum-yielding Taq DNA polymerase mutant controlled by stringent, multi-epitope antibody hot start
Supports efficient vortex mixing with proprietary anti-foaming formulation
SuperMix version provides maximum dye concentration for robust optical signal with small amplicons (i.e. microRNAtemplated cDNA)

PerfeCTa SYBR Green SuperMix is intended for molecular biology applications. This product is not intended for the diagnosis, prevention or treatment of a disease.

Description

PerfeCTa SYBR Green SuperMix is a user-friendly, 2X concentrated reaction mix that simplifies setup and is pre-blended with reference dye for optimized product performance. This proprietary buffer technology stabilizes a high concentration of SYBR Green I dye to ensure maximum optical signal with low abundance or small targets (such as microRNA). Successful detection with a non-specific, dsDNA intercalating dye requires precise target amplification as off-target primer elongation will contribute to overall fluorescent signal and lead to over-reported relative abundance values. This reagent is powered by a highly-processive, ultra-pure Taq DNA polymerase mutant with stringent, ultra-pure AccuStart™II antibody hot start technology that allows ambient room-temperature setup and maximal enzyme kinetics after rapid, irreversible denaturation at 95°C.

Performance Data

Comparison to QuantiTect, GAPD

Details

Contents
Single-tube, 2X concentrated reagent containing:
- Reaction buffer with optimized concentrations of molecular-grade MgCl2, dATP, dCTP, dGTP, and dTTP.
- AccuStart II Taq DNA Polymerase
- SYBR Green I dye
- Proprietary enzyme stabilizers and performance-enhancing additives.
- Titrated reference dye (if applicable).
Storage & Handling

Store components in a constant temperature freezer at -25°C to -15°C protected from light upon receipt. After thawing, mix thoroughly before using. 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®
- QIAGEN Rotor-Gene Q
No ROX
- Quantabio Q
- BioRad CFX
- Roche LightCycler 480
- Other
Bio-Rad iCycler iQ systems
- BioRad iCycler iQ™
- BioRad MyiQ™
- BioRad iQ™5
Related Resources

Product Manuals

PerfeCTa SYBR Green SuperMix for iQ PerfeCTa SYBR Green SuperMix PerfeCTa SYBR Green SuperMix, ROX PerfeCTa SYBR Green SuperMix, Low ROX

Product Flyers

PerfeCTa SYBR Green SuperMix/FastMix Flyer

Safety Data Sheets (SDS)

PerfeCTa SYBR Green SuperMix for iQ PerfeCTa SYBR Green SuperMix PerfeCTa SYBR Green SuperMix, ROX PerfeCTa SYBR Green SuperMix, Low ROX PerfeCTa SYBR Green SuperMix, UNG, for iQ PerfeCTa SYBR Green SuperMix, UNG PerfeCTa SYBR Green SuperMix, UNG, ROX PerfeCTa SYBR Green SuperMix, UNG, Low ROX

CofA (PSF)

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Publications

Increasing corn distillers solubles alters the liquid fraction of the ruminal microbiome
J. C.McCann, Journal of Animal Science - 2017
Abstract
Five ruminally fistulated steers were used in a 5 × 5 Latin square design to determine the effects of increasing dietary fat and sulfur from condensed distiller’s solubles (CDS) on the ruminal microbiome. Treatments included a corn-based control (CON) and 4 levels of CDS (0, 10, 19, and 27%) in a coproduct-based (corn gluten feed and soybean hulls) diet. Fat concentrations were 1.79, 4.43, 6.80, and 8.91% for diets containing 0, 10, 19, and 27% CDS, respectively. Steers were fed for ad libitum intake once daily. After feeding each diet for 18 d, ruminal samples were collected 3 h after feeding on d 19. Samples were separated into solid and liquid fractions. Microbial DNA was extracted for bacterial analysis using paired-end sequencing of the V3 through V4 region of the 16S rRNA gene on the MiSeq Illumina platform and quantitative PCR of selected species. Orthogonal contrasts were used to determine linear and quadratic effects of CDS inclusion. Increasing CDS inclusion decreased (linear, P < 0.05) α-diversity and species richness in the liquid fraction. Analysis of Bray–Curtis similarity indicated a treatment effect (P = 0.01) in the liquid fraction. At the phyla level, relative abundance of Bacteroidetes decreased in steers fed increasing dietary inclusion of CDS as Firmicutes increased to 82% of sequences for the 27% CDS treatment. Family Ruminococcaceae increased (linear, P < 0.01) 2-fold in the liquid fraction when feeding CDS increased from 0 to 27% CDS, yet genera Ruminococcus tended (P = 0.09) to decrease in steers fed greater CDS. The most abundant family of sulfate-reducing bacteria, Desulfovibrionaceae, increased (P < 0.03) in the solid and liquid fraction in steers fed additional dietary CDS and sulfur. Relative abundance of family Veillonellaceae and Selenomonas ruminantium were increased (linear, P ≤ 0.02) in the solid fraction as steers were fed increasing CDS. There were no effects (P > 0.10) of feeding increasing dietary fat from CDS on fibroylytic genus Fibrobacter in either fraction. Results demonstrate increasing fat and sulfur from CDS in a coproduct-based diet markedly alters the liquid fraction ruminal microbiome but does not elicit negative effects on relative abundance of identified fiber-fermenting bacteria.
YY1 Is Required for Posttranscriptional Stability of SOX2 and OCT4 Proteins
Mary C.Wallingford, Cellular Reprogramming - 2017
Abstract
Yinyang1 (YY1) participates in protein-DNA, protein-RNA, and protein–protein interactions and regulates developmental processes and disease mechanisms. YY1 interactions regulate a range of important biological functions, including oocyte maturation, epithelial to mesenchymal transition, and vascular endothelial growth factor (VEGF) signaling. We tested the hypothesis that YY1 is required for inner cell mass (ICM) lineage commitment during preimplantation development. In this study, we document gene expression patterns and protein localization of key transcription factors in Yy1 global, tissue-specific, and dsRNA-mediated knockout/down embryos. YY1 protein was found in cells of preimplantation and peri-implantation embryos, and adult tissues where two isoforms are observed. In the absence of YY1, OCT4 and SOX2 protein were lost in the ICM during preimplantation and naive neuroectoderm during gastrulation stages, yet no difference in Oct4 or Sox2 mRNA levels was observed. The loss of OCT4 and SOX2 protein occurred specifically in cells that normally express both OCT4 and SOX2 protein. These observations support a role for YY1 meditating and/or regulating the interaction of OCT4 and SOX2 at a posttranscriptional level. Our results suggest that distinct mechanisms of YY1-mediated molecular regulation are present in the early embryo, and may offer insight to promote lineage commitment in in vitro cell lines.
Curcumin Protects Skin against UVB-Induced Cytotoxicity via the
Maya Ben Yehuda Greenwald, Hindawi Oxidative Medicine and Cellular Longevity - 2017
Abstract
Curcumin was found to be beneficial in treating several skin pathologies and diseases, providing antioxidant protection due to its reducing properties and its electrophilic properties (the ability to activate the Nrf2 pathway and induce phase II cytoprotective enzymes). Nevertheless, clinical applications of curcumin are being hampered by its insufficient solubility, chemical instability,and poor absorption, leading to low efficacy in preventing skin pathologies. These limitations can be overcome by using a nanotechnology-based delivery system. Here, we elucidated the possibility of using curcumin encapsulated in a microemulsion preserving its unique chemical structure. We also examined whether curcumin microemulsion would reduce UVB-induced toxicity in skin. A significant curcumin concentration was found in the human skin dermis following topical application of a curcumin microemulsion. Moreover, curcumin microemulsion enhanced the reduction of UV-induced cytotoxicity in epidermal cells, paving the way for other incorporated electrophiles in encapsulated form protecting skin against stress-related diseases.
Bioaccessibility, bioavailability and anti-inflammatory effects of anthocyanins from purple root vegetables using mono- and co-culture cell models
Hua Zhang, Molecular Nutrition & Food Research - 2017
Abstract
Scope Immune-inflammatory, signalling and metabolic effects are the main pillars for bioactivity of anthocyanins derived from highly pigmented root vegetables. This study aims to assess the bioaccessibility and bioavailability of purple carrot and potato derived anthocyanins and the molecular mechanisms of their ability to ameliorate cellular inflammation in a mono- and co-culture cell models. Methods and Results An in vitro gastrointestinal model was used and demonstrated bioaccessibility of 44.62% and 71.8% for anthocyanins of purple carrot and potato, respectively. These accessible anthocyanins significantly inhibited cellular inflammation in Caco-2 cells. Intact cyanidin glycoside or petunidin glycoside (respectively from carrots and potatoes) were transported across a transmembrane cell model and detected by LC-MS/MS. Computational docking and glucose uptake analyses suggested uptake of anthocyanins was mediated by hexose transporters. Subsequent experiment using an inflamed Caco-2 BBe1/THP-1 co-culture cell model showed these transported anthocyanins inhibited IL-8 and TNF-α secretion, and expression of pro-inflammatory cytokines by blocking NF-κB, and MAPK mediated inflammatory cellular signalling cascades, but with varying degrees due to structural features. Conclusion Anthocyanins from purple carrots and potatoes possess a promising anti-inflammatory effect in model gut system. They can be absorbed and act differently but are in general beneficial for inflammation-mediated diseases. This article is protected by copyright. All rights reserved
Numerical Relationships Between Archaeal and Bacterial amoA Genes Vary by Icelandic Andosol Classes
Hendrikus J. Laanbroek, Microbial Ecology - 2017
Abstract
Bacterial amoA genes had not been detectable by qPCR in freshly sampled Icelandic Andosols thus far. Hence, a new primer set yielding shorter gene fragments has been designed to verify the absence of ammonia-oxidizing bacteria in different Icelandic Andosol classes. At the same time, a new primer set was also constructed for archaeal amoA genes that should improve the quality of PCR products. Although a large part of the soil samples were found to be amoA-negative, bacterial amoA genes were detectable with new as well as old primer sets. The same results were obtained for the archaeal amoA genes. The relative distribution of archaeal and bacterial amoA genes varied between Andosol classes. Archaeal amoA genes were significantly more abundant in Brown than in Histic Andosols, while the opposite was observed for bacterial amoA genes. The numbers of archaeal and bacterial amoA genes in Gleyic Andosols were not significantly different from those in Histic and Brown Andosols. The numbers of bacterial amoA genes, but not the numbers of archaeal amoA genes, correlated significantly and positively with potential ammonia oxidation activities. The presence of the bacterial nitrification inhibitor allylthiourea inhibited the potential ammonia oxidation activities during the first 12 h of incubation. Hence, it was concluded that ammonia-oxidizing bacteria profited most from the conditions during the measurements of potential ammonia oxidation activities.
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FAQs

Is there a SYBR® Green Fast Master Mix currently available
Yes. PerfeCTa SYBR Green Supermix is available. For further information, please consult the Kit protocol or product information.
What is the amount of cDNA used per TaqMan or SYBR qPCR Assay?
Suggested input quantities of template are: cDNA corresponding to 1 pg to 100 ng of total RNA; 100 pg to 100 ng genomic DNA. For more information, please consult the PerfeCta qPCR Supermixes or PerfeCta SYBR Green Supermixes protocols.
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Ordering Information

Product

Kit Size

Order Info

PerfeCTa SYBR Green SuperMix for iQ
Request Sample

100 x 50 μL rxns (2 x 1.25 mL)

Order (95053-100)

500 x 50 μL rxns (10 x 1.25 mL)

Order (95053-500)

2000 x 50 μL rxns (1 x 50 mL)

Order (95053-02K)
PerfeCTa SYBR Green SuperMix
Request Sample

100 x 50 μL rxns (2 x 1.25 mL)

Order (95054-100)

500 x 50 μL rxns (10 x 1.25 mL)

Order (95054-500)

2000 x 50 μL rxns (1 x 50 mL)

Order (95054-02K)
PerfeCTa SYBR Green SuperMix ROX
Request Sample

100 x 50 μL rxns (2 x 1.25 mL)

Order (95055-100)

500 x 50 μL rxns (10 x 1.25 mL)

Order (95055-500)

2000 x 50 μL rxns (1 x 50 mL)

Order (95055-02K)
PerfeCTa SYBR Green SuperMix Low ROX
Request Sample

100 x 50 μL rxns (2 x 1.25 mL)

Order (95056-100)

500 x 50 μL rxns (10 x 1.25 mL)

Order (95056-500)

2000 x 50 μL rxns (1 x 50 mL)

Order (95056-02K)

PerfeCTa SYBR® Green SuperMix