SensiMix™ SYBR® & Fluorescein Kit

Cat No. Size List Price* Qty  
QT615-05 500 Reactions
$980.00
 
QT615-20 2000 Reactions
$3,596.00
 
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*For more information on pricing please contact us

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SensiMix SYBR® & Fluorescein Kit is designed for superior sensitivity and specificity on real-time instruments that measure fluorescein signal as a passive reference for normalization. The mix has been developed to provide the high sensitivity required for demanding assays, or for those assays where template is limited.

Features & Benefits

  • Sensitivity - allows reproducible detection of low-copy number templates
  • Reproducibility - generates real-time data reliably and reproducibly
  • Specific - proprietary hot-start chemistry minimizes non-specific amplification
  • Flexible - for block-based instruments where fluorescein is required to allow calculation of dynamic well factors

Instrument Compatibility

Bio-Rad iCycler®, iQTM5 and MyiQ™. (See product selection table). However the kit can is also compatible with several instruments that do not require the use of fluorescein, such as the Qiagen Rotor-Gene™ 6000, the Bio-Rad CFX96 or the Roche LightCycler® 480.


Description

SensiMix SYBR® & Fluorescein Kit is a pre-mixed high performance product designed for superior sensitivity and specificity (Fig. 1) on various real-time instruments in which a Fluorescein signal is required. Powered by a proprietary, highly-optimized, chemical hot-start PCR enzyme, the mix has been developed to provide the high sensitivity and specificity required for demanding assays or for those assays where template is limited.

SensiMix SYBR® & Fluorescein is provided as a 2x mastermix containing all the components necessary for real-time PCR, including the SYBR® Green I dye, dNTPs, stabilisers and enhancers.

For expression analysis, SensiFAST cDNA Synthesis Kit is the perfect partner for SensiMix SYBR® & Fluorescein Kit for very high-quality first strand cDNA synthesis and real-time results.


Notes

SensiMix is a trademark of Bioline Reagents Ltd.


Components

Reagent

250 x 50µl Reactions

500 x 50µl Reactions

2000 x 50µl Reactions

SensiMix™ SYBR® & Fluorescein (2x)

5 x 1.25ml      (6.25ml)

10 x 1.25ml    (12.5ml)

40 x 1.25ml         (50ml)

50mM MgCl2

1 x 1ml

1 x 1ml

4 x 1ml


Volume

  • QT615-02: 250 x 50µl Reactions: 5 x 1.25ml
  • QT615-05: 500 x 50µl Reactions: 10 x 1.25ml
  • QT615-20: 2000 x 50µl Reactions: 40 x 1.25ml

Storage & Stability

All kit components should be stored at -20°C upon receipt for optimum stability. Repeated freeze/thaw cycles should be avoided. Avoid exposure of the SYBR® Green I to light.

When stored under the recommended conditions and handled correctly, full activity of the kit 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 SensiMix™ SYBR® & Fluorescein 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 17 COAs for SensiMix™ SYBR® & Fluorescein Kit →

Citations

  1. A signature of immune function genes associated with recurrence-free survival in breast cancer patients
    Ascierto M.L., Kmieciak M., Idowu M.O., Manjili R., Zhao Y., Grimes M., Dumur C., Wang E., Ramakrishnan V., Wang X.Y., Bear H.D., Marincola F.M., Manjili M.H.
  2. Induced Wnt5a expression perturbs embryonic outgrowth and intestinal elongation, but is well-tolerated in adult mice
    Bakker E.R., Raghoebir L., Franken P.F., Helvensteijn W., van Gurp L., Meijlink F., van der Valk M.A., Rottier R.J., Kuipers E.J., van Veelen W., Smits R.
  3. Human Monocytes Undergo Excessive Apoptosis following Temozolomide Activating the ATM/ATR Pathway While Dendritic Cells and Macrophages Are Resistant
    Bauer M., Goldstein M., Heylmann D., Kaina B.
  4. Anergic CD8+ T Lymphocytes Have Impaired NF-?B Activation with Defects in p65 Phosphorylation and Acetylation
    Clavijo P.E., Frauwirth K.A.
  5. Secreted Factors of Human Liver-Derived Mesenchymal Stem Cells Promote Liver Regeneration Early After Partial Hepatectomy
    Fouraschen S.M., Pan Q., de Ruiter P.E., Farid W.R., Kazemier G., Kwekkeboom J., Ijzermans J.N., Metselaar H.J., Tilanus H.W., de Jonge J., van der Laan L.J.

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 analysis 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 SensiMix SYBR® Kit for a probe assay?

      This is not possible because the buffer for SensiMix SYBR® kits and the buffer for probe kits are different, and have been optimised for their specific assays.
  • Why is the reaction volume 25µl or 50µl for this product?

      Most SensiMix reactions are 50µl with the exception of the SensiMix Capillary which is 20µl. These are the volumes recommended by the machine manufacturers, and can be scaled up or down if desired (for example when using a 384 well plate).
  • Why do the SensiMix 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 10 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.
  • Why are the ROX concentrations different?

      The amount of the ROX passive reference dye needed varies depending on the instrument optics, our SensiMix 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 Table), the SensiMix SYBR & Fluorescein contains fluorescein premixed in the mastermix at optimised concentrations.
  • Is the SYBR in a separate tube?

      For your convenience most of the SensiMix kits have an optimised amount of SYBR Green in the master mix. The exceptions are the SensiMix Capillary kits that have a separate vial of SYBR Green, so that it can be used for both probe-based and SYBR® Green-based chemistries on the capillary instruments, the SensiMix Probe Kits that do not require SYBR®.

  • 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?

      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 SensiMix 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

"SensiMix products from Bioline are easy to use and give consistent results, they make the switch to real-time PCR very simple."

Other researchers use:


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