SensiFAST™ Probe Hi-ROX Kit

Cat No. Size List Price* Qty  
BIO-82005 500 x 20µl Reactions
$412.00
 
BIO-82020 2000 x 20µl Reactions
$1,499.00
 
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*For more information on pricing please contact us

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SensiFAST™ Probe Hi-ROX Kit has been developed for fast, highly reproducible real-time PCR and has been validated on commonly used real-time PCR instruments.

A combination of the latest advances in buffer chemistry and PCR enhancers, together with a hot-start DNA polymerase, ensures that SensiFAST Probe Kit delivers fast, highly-specific and ultra-sensitive real-time PCR.

Features & Benefits

  • Highly specific - minimal non-specific activity leading to better efficiency and greater accuracy
  • Ultra-sensitive - perfect for difficult and low copy number samples
  • Efficient multiplexing - increasing throughput and conserving precious sample material
  • Fast - optimized for fast cycling conditions, ideal for high-throughput

Instrument Compatibility

ABI 7000, 7300, 7700, 7900, 7900HT, 7900HT FAST, StepOne™, StepOne™ Plus (see product selection table) each of these instruments having the capacity to analyze the real-time PCR data with the passive reference signal either on or off, as well as several instruments that do not require the use of ROX.


Description

SensiFAST™ Probe Hi-ROX Kit has been designed for fast, highly reproducible real-time PCR and has been validated on commonly used real-time instruments. The kit is formulated for use with probe-detection technology, including TaqMan®, Scorpions® and molecular beacon probes. SensiFAST Probe Kit is optimized for fast mode on fast qPCR instruments and fast cycling conditions on standard qPCR instruments.

A combination of the latest advances in buffer chemistry and PCR enhancers, together with a hot-start DNA polymerase, ensure that SensiFAST Probe Kit delivers shorter run times with results that are highly reproducible, highly-specific and ultra-sensitive (Fig. 1), making SensiFAST ideal for high-throughput assays. The advanced buffer chemistry and enhancers make SensiFAST Probe perfect for multiplexing (Fig. 2).

For expression analysis, SensiFAST cDNA Synthesis Kit is the perfect partner for SensiFAST Probe Hi-ROX Kit for very fast cDNA synthesis and real-time results, without compromising sensitivity or quality.

SensiFAST Probe Hi-ROX is provided as a 2x mastermix containing all the components necessary for real-time PCR, including dNTPs, stabilizers and enhancers. SensiFAST Probe Hi-ROX contains premixed ROX for optional use.


Notes

SensiFAST is a trademark of Bioline Reagents Ltd.


Components

Reagent

500 x 20 µL reactions

2000 x 20 µL reactions

SensiFAST Probe Hi-ROX mix (2x)

5 x 1 mL

4 x 5 mL

 


Volume

  • BIO-82002: 200 x 20µl Reactions: 2 x 1ml
  • BIO-82005: 500 x 20µl Reactions: 5 x 1ml
  • BIO-82020: 2000 x 20µl Reactions: 4 x 5ml

Storage & Stability

All kit components should be stored at -20°C upon receipt. When stored under the recommended conditions and handled correctly, full activity of the kit is retained until the expiry date on the outer box label. Avoid exposure of the ROX™ to light.


Shipping conditions

Shipped on dry/blue ice.


Certificates of Analysis (COAs)

Certificates of Analysis for SensiFAST™ Probe Hi-ROX Kit are grouped by catalogue number and then listed by batch number. Larger pack sizes are listed first. To locate the COA for your product, first find the catalogue number and then the required batch number.

View all 35 COAs for SensiFAST™ Probe Hi-ROX Kit →

Citations

  1. The ?-1, 4-endoglucanase gene is suitable for the molecular quantification of the root-lesion nematode, Pratylenchus thornei
    Mokrini, F., Waeyenberge, L., Viaene, N., Andaloussi, F.A., Moens, M.
  2. Tumor Progression Locus 2 Differentially Regulates IFN? and IL-17 Production by Effector CD4+ T Cells in a T Cell Transfer Model of Colitis
    Acuff, N.V., Li, X., Kirkland, R., Nagy, T., Watford, W.T.
  3. Depletion of 14-3-3? reduces the surface expression of Transient Receptor Potential Melastatin 4b (TRPM4b) Channels and attenuates TRPM4b-mediated glutamate-induced neuronal cell death
    Cho, C.H., Kim, E., Lee, Y.S., Yarishkin, O., Yoo, J.C., Park, J.Y., Hong, S.G., Hwang, E.M.
  4. Conserved sequence-specific lincRNA–steroid receptor interactions drive transcriptional repression and direct cell fate
    Hudson, W.H., Pickard, M.R., de Vera, I.M.S., Kuiper, E.G., Mourtada-Maarabouni, M., Conn, G.L., Kojetin, D.J., Williams, G.T., Ortlund, E.A.
  5. Essential role of Elmo1 in Dock2-dependent lymphocyte migration
    Stevenson, C., de la Rosa, G., Anderson, C.S., Murphy, P.S., Capece, T., Kim, M., Elliott, M.R.

Frequently asked questions

  • What is the advantage of working with SYBR Green I?

      SYBR Green I is an inexpensive, universal dye which binds to all dsDNA. It can be easily used in combination with a simple primer pair to detect PCR products in real-time. This dye is very attractive for researchers analysing lots of different genes.

  • What is the advantage of working with a probe system?

      The probe system is always specific; only detect the gene of interest. It is also possible to distinguish between similar sequences with small differences like SNPs or mutations. In general, probe assays need less optimisation than SYBR Green I assays.

  • Can I use a SensiFAST Kit for standard real-time PCR?

      Although SensiFAST kits have been designed for fast PCR on the new generation of fast machines, they will work equally well for standard or fast PCR protocols on all real-time PCR machines.
  • Can I use a SensiFAST SYBR Kit for a probe assay?

      This is not possible because the SYBR is pre-mixed into the SensiFAST SYBR mastermix.
  • Why do the SensiFAST kits contain a hot-start Taq polymerase?

      Polymerase activity during the reaction set-up causes non-specific amplification including primer-dimer formation. To avoid non-specific amplification the polymerase is only activated after heating at 95°C for 2-3 minutes.
  • Why do you no longer sell kits contain dUTP (and UNG)?

      This is because it is because it is only effective if all the researchers either doing PCR in the laboratory or using the same thermocycler are also using a dUTP/dNTP and UNG system. If even one researcher is not, it ceases to be an effective control. Using dUTP has also been shown to be inefficient as it increases the Ct values by reducing reaction efficiency. Most labs do not need to use the dUTP method of control, optimised protocols will allow high specificity of PCR and good lab practices (using disposable consumables, the use of filter tips and maintaining a separate area for PCR set-up and PCR amplification and any post-PCR analysis) virtually eliminate the risk of cross contamination.
  • Why do certain kits contain a ROX passive reference?

      The emission recorded from ROX during the baseline cycles is used to normalize the emission recorded from the reporter (SYBR) in later cycles in some instruments. ROX compensates for small fluorescent fluctuations such as bubbles and well-to-well variations that may occur. Each of these instruments having the capacity to analyze the real-time PCR data with the passive reference signal either on or off.
  • Why are the ROX concentrations different?

      The amount of the ROX passive reference dye needed varies depending on the instrument optics, our SensiFAST kits have been optimised for these different instruments (see Product Selection Tool).
  • What is the difference between using ROX and using fluorescein?

      Fluorescein is an alternative passive reference dye used just in the Bio-Rad instruments (see Product Selection Tool), the SensiFAST SYBR & Fluorescein contains fluorescein premixed in the mastermix at optimised concentrations.
  • Is the SYBR in a separate tube?

      For your convenience the SensiFAST kits have an optimised amount of SYBR Green in the mastermix.
  • What is the difference between a one-step and a two-step real-time PCR reaction?

      In a one-step reaction the reverse transcription reaction and the real-time PCR reaction are done in one tube, making this a closed tube assay, so contamination can be avoided. It saves pipeting steps and time, and is easy in handling, making it ideal for high throughput screening.

      In a two-step reaction the reverse transcription reaction and the real-time PCR reaction are done in separate tubes. It gives a more flexible way of working in that the cDNA can be used for more than one real-time PCR reaction and can be archived, eliminating the need to continually isolate RNA. For convenience Bioline sells the SensiFAST cDNA Synthesis Kit separately if you wish to take a two-step approach.
  • What template (RNA/cDNA) is this compatible with?

      SensiFAST One-Step Kits can be used with most RNA/cDNA templates. To give you some idea of a few of the types of templates used with these kits:

      Human - Rosato R.R., et al. Cancer Res. 67: 9490-500 (2007)
      Rat - Remund K., et al. Expt. Lung Res. 35: 359-370 (2009)
      Drosophila - Brown A.E., et al. PloS ONE 4: e4490 (2009)
      Nitrogen-fixing bacteria - Bahlawane C., et al. Mol. Plant-Microbe Inter. 21: 1498-1509 (2008)
      Diphtheria bacteria - Jochmann N., et al. Microbiology 155: 1459-1477 (2009)
      Green algae - Wobbe L., et al. PNAS 106: 13290-13295 (2009)
      Bovine virus - Park S-I., et al. J. Virol. Methods. 159: 64-6 (2009)

      For cDNA/DNA templates SensiFAST SYBR and Probe kits are used. To give you some idea of a few of the types of templates used with these kits:

      Stem Cell - Bernardo A.S., et al. Stem Cells 27(2): 341 -351 (2008)
      Cow - Baumert A., et al. J. Dairy Research 76(3): 356-364 (2009)
      Mouse - Hoyles R.K., et al. Arthritis Rheum. 58(4): 1175-88 (2008)
      Quail - De Winter P., et al. British Poultry Science 49(5): 566 – 573 (2008)
      Insect - Bass C., et al. Malaria journal 6 111 (2007)
      Fish - Miller M.R., et al. J. Nutr. 138(11): 2179-2185 (2008)
      Crab - Wilcockson D.C. & Webster S.G. Gen. & Comp. Endo. 156: 113–125 (2008)
      Nematode - Murray S.L., et al. Mol. Plant-Microbe Interactions 20(11): 1431–1438 (2007)
      Plant - Hecht V., et al. Plant Physiology Preview DOI:10.1104/pp.107.096818 (2007)
      Yeast - Kawauchi J., et al. Genes & Dev. 22: 1082-1092 (2008)
      E.coli Bacteria - Saeed H.A., et al. Research J. Microbiology 4(4); 173-177 (2009)
      Virus - Muscillo M., et al. Water, Air, & Soil Pollution 191: 1-4 (2008)


Customer Testimonials

Our lab has been routinely been using the Bioline SensiFAST Probe Hi-ROX for microRNAs detection in mammalian cells for the past 3 years. Results are highly reproducible between technical replicates and independent experiments. We found that this product showed high sensitivity, low background noise and consistent amplification curves. It performs as well if not better than competitor products. In addition, several thawing/freezing cycles do not influence its activity.
Given its low cost, SensiFAST Probe Hi-ROX is a really attractive option compared to competitor products. Overall, we are completely satisfied and would recommend it to other labs.
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"I have tried multiple master mixes to optimize my qPCR with complex environmental sample. With Bioline's SensiFAST I was able to dramatically boost my efficiencies and increase reproducibility. In addition, I found that multiple primer pairs produced cleaner products with improved melting curves."

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