SensiFAST™ cDNA Synthesis Kit

Cat No. Size List Price* Qty  
BIO-65053 50 Reactions
$307.00
 
BIO-65054 250 Reactions
$1,347.00
 
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SensiFAST™ cDNA Synthesis Kit has been developed to generate high-quality first strand cDNA from total RNA template. The kit contains all necessary components in a unique TransAmp™ buffer system to deliver the highest quality cDNA.

Features & Benefits

  • Reproducible – unique reverse transcriptase and new TransAmp™ buffer generate consistent first-strand cDNA
  • Broad dynamic range – ideal for dilute and low-copy samples
  • Sensitive – real-time RT-PCR analysis from as little as 1pg of starting total RNA
  • Unbiased cDNA synthesis – complete 5’ to 3’ RNA sequence representation
  • Rapid – cDNA synthesis and real-time PCR in less than 60 minutes, ideal for high throughput applications

Applications

  • Two-step RT-PCR
  • Two-step RT-qPCR

Description

SensiFAST cDNA Synthesis Kit provides a rapid and sensitive method for first-strand cDNA synthesis for use in real-time PCR studies (Fig. 1). SensiFAST cDNA Synthesis Kit displays excellent linearity across a wide range of starting material, revealing the same relative representation in your cDNA template, regardless of gene abundance. Furthermore, SensiFAST cDNA Synthesis Kit will produce reliable, high performance results, in both real-time and conventional RT-PCR.

A reverse transcriptase and new TransAmp buffer system delivers both highly efficient first-strand synthesis and higher cDNA yields, leading to enhanced reproducibility (Fig. 2) and data accuracy, making SensiFAST cDNA Synthesis Kit ideal for working with limited samples, such as laser-microdissected samples and tissue biopsies. The TransAmp Buffer also employs a unique blend of random hexamer primers and anchored oligo dT to ensure unbiased 3' and 5' coverage and reverse transcription of all regions (Fig. 3). Additionally SensiFAST cDNA Synthesis Kit can be used with SensiFAST Probe and SYBR® Kits for very fast cDNA synthesis without compromising quality (Fig. 4), with real-time results in less than an hour.


Notes

SensiFAST and TransAMP are trademarks of Bioline.


Components

 Product Name

 50 reactions

 250 reactions

 5x TransAmp Buffer

 200µl

 1ml

 Reverse Transcriptase

 50µl

 250µl


Storage & Stability

SensiFAST cDNA Synthesis Kit can be stored for up to 12 months at -20°C. Repeated freeze/thaw cycles should be avoided.

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


Shipping conditions

On dry or blue ice.


Certificates of Analysis (COAs)

Certificates of Analysis for SensiFAST™ cDNA Synthesis 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 26 COAs for SensiFAST™ cDNA Synthesis Kit →

Citations

  1. Generation and characterization of small single domain antibodies inhibiting human tumor necrosis factor receptor 1
    Steeland, S., Puimège, L., Vandenbroucke, R.E., Van Hauwermeiren, F., Haustraete, J., Devoogdt, N.,Hulpiau, P., Leroux-Roels, G., Laukens, D., Meuleman, P., De Vos, M., Libert, C
  2. BRAF-and MEK-Targeted Small Molecule Inhibitors Exert Enhanced Antimelanoma Effects in Combination With Oncolytic Reovirus Through ER Stress
    Roulstone, V., Pedersen, M., Kyula, J., Mansfield, D., Khan, A.A., McEntee, G., Wilkinson, M., Karapanagiotou, E., Coffey, M., Marais, R., Jebar, A., Errington-Mais, F., Melcher, A., Vile, R., Pandha, H., McLaughlin, M., Harrington, K.J.
  3. Regulation of Hepatocyte Growth Factor in Mice with Pneumonia by Peptidases and Trans-Alveolar Flux
    Raymond, W.W., Xu, X., Nimishakavi, S., Le, C., McDonald, D.M., Caughey, G.H.

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

The SensiFast cDNA Synthesis Kit was directly compared to another cDNA synthesis kit. During real-time qPCR, the CT values of the cDNA samples produced with the SensiFast cDNA synthesis Kit lay between 16 and 18. The samples behaved very consistent and results were highly reproducible. The cDNA samples produced with the other kit had CT values between 24 and 28 and were much more scattered.
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I have been using Bioline products for past 5 years, starting with MyTaqRed Mix and dNTPs. Recently, we started using the SensiFAST cDNA Synthesis Kit and SensiFAST SYBR mix for qRT-PCR. Upon testing with diverse RNA sources, we were really satisfied with the results as the data is quite comparable with competitor SYBR mixes. Quality to price ratio is the winning factor for both the cDNA synthesis kit and SYBR mix. We felt real ease-of-use with the cDNA synthesis kit, which was fast and simple. I recommend the SensiFAST cDNA Synthesis kit and SensiFAST SYBR mix from Bioline.
Working with post-mortem human heart tissue samples for a gene expression project proved to be a challenging task for tissue samples that came from the medical examiner's office. So many factors were working against a viable gene expression profile for these samples such as time of death to autopsy lag, time from autopsy to tissue sample, and whether or not the tissue had been flash frozen. The samples were also never treated with RNA-later so we expected highly degraded RNA from these tissue samples.

Our only light of hope was the fact that our target amplicon sizes were less than 100 bp which meant that we could still find what we were looking for in spite of highly degraded RNA. Chances were that at least a few intact sequences would amplify. The critical part in our experiment was to be able to yield enough cDNA from whatever bit of RNA we were able to extract from our samples.

This is where Bioline exceeded all expectations. Working quickly and carefully, I was able to get decent cDNA considering all the factors working against my RNA isolations. The cDNA was enough to be able to carry out a gene expression profile for each of the samples and our project was a success. Bioline proved to be superior in cDNA yield from highly degraded RNA samples.
I had been repeating an experiment over and over again because I wasnt getting any amplicon of my target gene after cDNA synthesis and qPCR, thinking my differentiation protocol wasnt working until I tried the SensiFAST cDNA Synthesis Kit and bammmmm, the amplicon was there. I was using more than 50 ng/µl with a different company and wasn't getting any result but with just 10 ng/µl on the SensiFAST cDNA Synthesis Kit, I get to save some RNA for future reference and get the data I am looking for. Are you working with lesser concentration of RNA that you need to be able to replicate your experiments? SensiFAST cDNA Kit is for you then!
"My aim is to isolate as much RNA as possible from a small amount of primary cells, which then are used for further qPCR experiments.
With the help of Bioline products (ISOLATE II RNA Micro Kit and SensiFAST cDNA Synthesis Kit) it is possible to achieve the best results with the lowest amount of sample."

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