SensiFAST™ Probe Hi-ROX One-Step Kit

Cat No. Size List Price* Qty  
BIO-77001 100 x 20µl Reactions
$215.00
 
BIO-77005 500 x 20µl Reactions
$927.00
 
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*For more information on pricing please contact us

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SensiFAST™ Probe Hi-ROX One-Step Kit has been specially formulated for highly reproducible first-strand cDNA synthesis and subsequent real-time PCR in a single tube.

The kit is designed for use with probe-detection technology. It uses the latest advances in buffer chemistry, together with a reverse transcriptase and hot-start DNA polymerase system to produce fast, highly-specific and ultra-sensitive one-step real-time RT-PCR.

Features & Benefits

  • Accurate quantification - perfect for calculating relative gene expression from RNA samples
  • Ultra-sensitive - unique buffer chemistry gives superior sensitivity of low copy targets
  • Rapid - optimized proprietary enyzme and buffer chemistry for fast cycling
  • Flexible - compatible with all standard and fast cycling instruments

Instrument Compatibility

ABI 7000, 7300, 7700, 7900, 7900HT, 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 One-Step Kit has been optimized for fast mode on fast real-time RT-PCR instruments and fast cycling conditions on standard real-time PCR instruments. The kit is designed for superior sensitivity and specificity with probe-detection technology, including TaqMan®, Scorpions®and molecular beacon probes.

SensiFAST Probe Hi-ROX One-Step Kit has been formulated for highly reproducible first-strand cDNA synthesis and subsequent real-time PCR in a single tube. The antibody-mediated hot-start DNA polymerase system reduces the chances of primer/dimers formation. This allows a much greater dynamic range by removing competition for reaction components during amplification, in turn leading to greater sensitivity (Fig. 1). The advanced buffer chemistry and enhancers also give SensiFAST Probe Hi-ROX One-Step unbeatable efficiency in multiplexing (Fig. 2). SensiFAST Probe Hi-ROX One-Step Kit consists of a 2x SensiFAST Probe One-Step mix, separate reverse transcriptase and RiboSafe RNase Inhibitor.

SensiFAST Probe Hi-ROX One-Step Kit also contains pre-mixed ROX for optional use.


Notes

SensiFAST is a trademark of Bioline.


Components

Reagent

100 x 20µl Reactions

500 x 20µl Reactions

SensiFAST Probe Hi-ROX One-Step mix (2x)

1 x 1ml

5 x 1ml

RiboSafe RNase Inhibitor

1 x 40µl

1 x 200µl

Reverse transcriptase

1 x 20µl

1 x 100µl

DEPC-H2O

1 x 1.8ml

2 x 1.8ml


Volume

  • BIO-77001: 100 x 20μl Reactions: 1 x 1ml
  • BIO-77005: 500 x 20μl Reactions: 5 x 1ml

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

SensiFAST Probe Hi-ROX One-Step Kit is shipped on dry/blue ice


Certificates of Analysis (COAs)

Certificates of Analysis for SensiFAST™ Probe Hi-ROX One-Step 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 25 COAs for SensiFAST™ Probe Hi-ROX One-Step Kit →

Citations

  1. Involvement of YAP, TAZ and HSP90 in Contact Guidance and Intercellular Junction Formation in Corneal Epithelial Cells
    Raghunathan, V.K., Dreier, B., Morgan, J.T., Tuyen, B.C., Rose, B.W., Reilly, C.M., Murphy, C.J.
  2. Development of a surveillance scheme for equine influenza in the UK and characterisation of viruses isolated in Europe, Dubai and the USA from 2010–2012
    Woodward, A.L., Rash, A.S., Blinman, D., Bowman, S., Chambers, T.M., Daly, J.M., Damiani, A., Joseph, S., Lewis, N., McCauley, J.W., Medcalf, L., Mumford, J., Newton, J.R., Tiwari, A., Bryant, N.A., Elton, D.M.
  3. Human coronavirus NL63 replication is cyclophilin A-dependent and inhibited by non-immunosuppressive cyclosporine A-derivatives including Alisporivir
    Carbajo-Lozoya, J., Ma-Lauer, Y., Maleševi?, M., Theuerkorn, M., Kahlert, V., Prell, E., von Brunn, B., Muth, D., Baumert, T.F., Drosten, C., Fischer, G., von Brunn, A.

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)



Other researchers use:


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