SensiFAST™ Hi-ROX Genotyping Kit

Cat No. Size List Price* Qty  
BIO-35002 200 x 20µl Reactions
$161.00
 
BIO-35005 500 x 20µl Reactions
$416.00
 
BIO-35020 2000 x 20µl Reactions
$1,512.00
 
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SensiFAST™ Hi-ROX Genotyping Kit is designed for fast, precise and highly reproducible genotyping. A combination of the latest advances in buffer chemistry, together with an antibody-mediated hot-start DNA polymerase system, ensure that the SensiFAST Hi-ROX Genotyping Kit produces highly-specific, ultra-sensitive real-time PCR with clear allelic discrimination and outstanding allele clustering.

Features & Benefits

  • Accurate allele calling - clear allele discrimination for higher confidence genotype calling
  • Tight fluorescence clusters - demonstrate high reproducibility and are perfect for difficult SNPs
  • Faster run time - enabling more experiments per day, ideal for large genotyping projects
  • Designed for dual-labeled probe assays - for example TaqMan® probe-based assays

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 Hi-ROX Genotyping Kit has been developed for fast, precise and highly reproducible genotyping of sequence variants, including loci with type IV SNPs. The SensiFAST Hi-ROX Genotyping Kit is a combination of the latest advances in buffer chemistry, together with an antibody-mediated hot-start DNA polymerase system. This produces highly-specific, ultra-sensitive real-time PCR with clear allelic discrimination and outstanding allele clustering (Fig. 1). The SensiFAST Hi-ROX Genotyping Kit is compatible with many dual-labeled probe assays (including TaqMan® probe based assays) and has been validated on commonly used real-time instruments.

The advanced buffer chemistry in the SensiFAST Hi-ROX Genotyping Kit provides higher stringency and more specific binding of the allele-specific probes, resulting in much narrower probe melting temperature windows. The combination of these developments leads to wider and clearer separation of allele clusters. The kit allows robust genotype calling, enabling reproducible and accurate results. The advanced buffer also means that minimal optimization is required to achieve fast, reproducible and accurate results, ideal for high-throughput in large-scale genotyping projects.


Notes

SensiFAST is a trademark of Bioline Reagents Ltd.


Components

Reagent

200 x 20µl Reactions

500 x 20µl Reactions

2,000 x 20µl Reactions

SensiFAST Hi-ROX Genotyping Mix (2x)

2 x 1ml

5 x 1ml

20 x 1ml


Volume

  • BIO-35002: 200 x 20µl Reactions: 2 x 1ml
  • BIO-35005: 500 x 20µl Reactions: 5 x 1ml
  • BIO-35020: 2,000 x 20µl Reactions: 20 x 1ml

Storage & Stability

All components should be stored at -20°C upon receipt for optimum stability. Repeated freeze/thaw cycles should be avoided.

When stored under the recommended conditions and handled correctly, full activity of the reagents is retained until the expiry date indicated on the outer box label.


Shipping conditions

Shipped on dry/blue ice.


Certificates of Analysis (COAs)

Certificates of Analysis for SensiFAST™ Hi-ROX Genotyping 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 6 COAs for SensiFAST™ Hi-ROX Genotyping Kit →


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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