Real-time quantitative PCR (qPCR) analysis
TATAA Biocenter offers to perform your qPCR analyses. Our experts pioneered qPCR based nucleic acid analyses in Europe and have more than a decade of experience of projects and research within the field. Our facilities hold a unique instrument park with a broad selection of qPCR instruments from all leading manufacturers, including fast cyclers, High Resolution Melt (HRM) cyclers, and high throughput instruments. Applications such as gene expression analysis, genotyping, SNP analysis, HRM, Digital PCR , immuno-qPCR , and any more esoteric application of your choice are easily performed with confidence. TATAA’s experts can also help you select the optimum reference genes for your analyses.
Do you want to perform superior gene expression analysis?
TATAA Biocenter is a proud partner of Life Technologies TaqMan® Service Provider Program.
Examples of qPCR analyses that are performed at TATAA Biocenter:
- Gene expression analysis (relative and absolute quantification)
- Copy number variation (CNV) analysis
- miRNA analysis
- DNA methylation analysis
- CpG-methylation analysis
- SNP typing
- Allelic discrimination (end-point analysis)
- Digital PCR
- Protein quantification with immuno qPCR
- Pathogen detection and quantification
- qPCR data analysis and data mining with professional softwares
- Optimum reference gene analysis
or other applications of your choice.
Either send your prepared DNA or cDNA to us for real-time PCR analysis, or combine your qPCR with our other services e.g. nucleic acid extraction, experimental design, assay design and validation, advanced analysis etc. for a complete workflow.
High-throughput qPCR analysis
TATAA Biocenter is meeting the challenges of today’s high-throughput research environment by offering exceptionally cost efficient high-throughput analysis services on the Life Technologie’s QuantStudio™ 12K Flex OpenArray® Platform and the BioMark™ system from Fluidigm.
The QuantStudio™ 12K Flex OpenArray® Platform (placed at TATAA Gothenburg) is a high-throughput qPCR instrument with four main applications: Gene Expression, Digital PCR, miRNA profiling, and Genotyping. OpenArray plates consist of 3072 through-holes that can be loaded with reagents to perform individual 33 nl qPCR reactions. Four plates can be processed per run, which gives 12 288 reactions/run, which is equal to 128 runs on a 96-well based instrument.
Choose between using your own custom TaqMan® assays, inventoried TaqMan® assays, or use the pre-designed Pathway Panels.
For more information about the pre-designed Pathway Panels from Life Technologies and to see the complete gene list for these panels please visit here.
The BioMark™ system (placed at TATAA Gothenburg and at our Prague facilities) is a high-throughput qPCR instrument that uses integrated fluidic circuits known as dynamic and digital arrays for the applications Digital PCR, Gene Expression, and Genotyping. The dynamic arrays are suited for either 48 samples and 48 assays (2304 reactions) or 96 samples and 96 assays (9216 reactions). The digital arrays has 12.765 chips (for 12 samples and 765 reactions/sample) and 48.770 chips (for 48 samples and 770 reactions/sample).
This innovative instrument allows practice of tried-and-true techniques, such as TaqMan® assays, while realizing a previously unachievable throughput. Researchers save on reagents, pipette tips and time. Far more data are generated compared to microwell chips, and with no pre-spotting required.
Use as little as 10 picograms of starting material – reproducible results from populations of cells down to a single cell.
Optimum reference gene analysis
The best way to choose the proper reference gene for your present sample setup is by running a panel of potential genes on a number of representative test samples. The gene(s) most appropriate for normalization are chosen in each case by analyzing the data with the powerful tool GenEx for statistical qPCR data processing and analysis. TATAA Biocenter offers to analyze your samples and choosing the most appropriate reference gene, either by running genes from our Human, Mouse or Rat Reference Gene Panel with commonly used reference genes in gene expression studies. The genes have varying cellular function and expression levels. Or we can run your in-house genes, alternatively design and validate the genes of your choice, also for other organisms than human or mouse.
For all gene expression studies using quantitative PCR it is necessary to compensate for differences between samples due to material losses, differences in reverse transcription effiencies and PCR inhibition. Normalization should include measuring a reference gene, and can also be complemented by identical sample-input amounts. The reference gene should have a constant expression in all the samples compared in your present study. However, there is no universal control gene that can be used for all different studies, expressed at a constant level under all conditions and in all tissues.
Digital PCR is an exceptionally accurate and sensitive method to detect and count individual nucleic acid molecules for applications such as detection of rare mutations, quantifying viral load, gDNA, cDNA, plasmids, or next-generation sequencing libraries.
TATAA Biocenter offers Digital PCR services on the Life Technologies OpenArray® system (placed at TATAA Gothenburg) and the BioMark™ system from Fluidigm (placed at our Prague facilities). It also possible to visit us for hands-on training to learn the technique. Digital PCR is based on the concept that PCR produces a very large amount of product even from a single template molecule. In order to determine the number of molecules in a sample with high precision, the sample is divided into a large number of aliquots/reactions that are analyzed separately by traditional end-point PCR. Some of these reactions contain the target molecule (positive) while others do not (negative). Hence, by calculating the number of positive PCR’s, virtually the number of template molecules that were present initially in the sample is counted, without reference to standards or endogenous controls.
With Digital PCR it is convenient to run many reactions fast. The OpenArray® plates consist of 3072 through-holes (reaction chambers) per plate, and three plates can be processed per run. The BioMark™ system has two different digital arrays, the 12.765 chips (for 12 samples and 765 reactions/sample) and the 48.770 chips (48 samples and 770 reactions/sample).
At the following link (will open YouTube in a new window) you will find a video describing the technology: Life Technologies “Digital PCR – Absolute Quantification for your science!”.
Read more about Digital PCR:
Single cell analysis
TATAA has many years of experience of single cell analysis with qPCR. Single cell gene expression profiling is a powerful tool to study heterogeneity between and within cell types/populations. Furthermore, it allows screening for rare cells, such as stem cells.
Single cell gene expression profiling has traditionally been technically challenging since the total number of transcript is low. Therefor we have developed CelluLyser, which is a lysis buffer that minimizes RNA/DNA losses and is compatible with downstream RT-qPCR applications.
We have experience of analyzing different types of cells, such as embryonic stem cells, astrocytes and beta cells. Using a validated workflow, we collect, lyse, reverse transcribe, and measure transcript with qPCR using carefully optimized assays. If several targets are to be analyzed we pre-amplify our targets. We have also developed statistical tools to characterize single cells and identify new cell types.
Sensitive protein analysis with immuno-qPCR
TATAA Biocenter offers quantification of proteins by immuno-qPCR, making it possible to study your proteins with high sensitivity using real-time PCR. Combining the latest technologies in the field and many years of experience, your samples will be professionally analyzed by our experts. Immuno-qPCR has a broader dynamic range than ELISA, from low to high concentrations of antigen, with the possibility to increase the sensitivity of your analyses 100-1000 times compared to ELISA. Different DNA labels make it possible to multiplex your immuno-qPCR assays. Immuno-qPCR are especially useful for lowly expressed disease markers, low load infections, small samples volumes etc.
- Conjugation of DNA to antibody of your choice and purification
- Full immuno-qPCR assay development and optimization
- Full range immuno-qPCR projects including analysis
- Immuno-qPCR hands-on courses
TATAA Biocenter offers plasmid quantification services, using real-time PCR. Simply provide the sample material and we will design assays optimized for your expression system. Use our plasmid copy number (PCN) analyses to control your protein production on the plasmid level, to follow the plasmid copy number changes over time, for quality control of the material from your cell banks, or to check purity after changing production systems, etc.
TATAA Biocenter has many years of experience in developing assays for pathogen detection, both for food diagnostic and clinical applications. TATAA has also extensive experience in sample preparation from complex matrices which is often a limiting step in pathogen detection.
Another important aspect is training. Pathogen detection is often performed by microbiologists and their experience in molecular biological techniques is sometimes limited. Therefore, when implementing a new qPCR based method it can be important to train the staff properly on the new method. As one of TATAA’s main field of services is training, we are the perfect match also in these cases.
TATAA can today offer a full range of services for qPCR based pathogen detection including assay design for detection and subtyping, development of extraction methods, data analysis, validation of methods and training. We do of course also perform routine analysis of existing methods or of the method we develop specifically for you.
Expression analysis of circulating tumour cells (CTC)
TATAA Biocenter can offer expanded detection and analysis services where AdnaGen kits are used for the CTC isolation. The detection is performed by TATAA using custom designed probes and assays based on the customers need.
The AdnaGen system for isolation and detection of CTC is called AdnaTest and is presently CE-certified for diagnostic use for breast, colon and prostate cancer within Europe. The Company’s tumor diagnostics are complete kits consisting of reagents for immunomagnetic selection, mRNA isolation and multiplex RT-PCR, and can be performed on standard technology platforms. The products have high analytical sensitivity (2 cells in 5 ml blood) and high specificity (>90%) achieved through the combination of multi-marker tumor cell enrichment and multiplex gene expression profiling.
All the AdnaTest kits are standardized and use a predetermined set of markers to detect CTC. These markers have all been carefully selected to maximize the probability of finding CTC in a sample and there is a growing body of scientific literature and publications based on these kits, which makes comparisons between studies relatively easy.
AdnaTest kits are also available in TATAA Biocenter’s Product page. For more information about the kits or to make an order, visit Products – CTC tests
At TATAA Biocenter we perform post-PCR work in separated areas to avoid contamination risks. We purify PCR product for downstream work or confirm the specificity of assays by gel electrophoresis.
or other projects of your choice
For other projects of your choice, or the mentioned above, please feel free to contact us for requests and more information .