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Digital PCR Analysis

 
 
Digital PCR has become the method of choice to provide precise absolute quantification of nucleic acids. The big advantage over conventional qPCR approaches is that digital PCR does not require any normalization to controls (either to a reference or to a standard curve).
 

Features and Benefits

 
High Quality
  • Digital PCR assays with highest accuracy
  • High sensitivity due to optimization of PCR working conditions: we design, synthesize and test your primer/probe set(s) for their PCR performance

 

Expert Knowledge
  • Merging together our experience in sample handling, assay design and optimization as well as validation we perform analysis of your samples by digital PCR with the highest efficiency

 

 
Convenient
  • Synthesis of primers for use in EvaGreen assays
  • Synthesis of primers and qPCR probes which are compatible with commercially available supermixes
  • Availability of various TM enhancers (LNA, MGB, pdC/pdU) to   
    • To improve specificity and accuracy of drop-off assays (duplexed primer probe assays with a drop-off as well as a reference probe) for detection of NHEJ-induced CRISPR/Cas9 mutations
    • For optimizing the annealing temperature of primers to be used in ddPCR in order to reduce “rain” artifacts*
  • Synthesis of high-purity probes to increase your confidence when using ddPCR as molecular detection platform for liquid biopsy
  • Design, optimization, validation and analysis from one source
  • Can be combined with other Microsynth services (e.g. nucleic acid isolation)
 
 
* Digital droplet PCR works on the premise of dividing a sample into small compartments or droplets, each containing a single molecule of interest. The droplet's fluorescence is examined. If the fluorescence reaches a cutoff the droplet is then defined as positive, if not it is defined as negative. The concentration within the sample as a whole is then calculated using the proportion of positive and negative droplets and Poisson statistics. Droplets that are not clearly defined as negative or positive are called rain droplets.
 

Applications

 
Currently Microsynth is offering a digital PCR analysis service for R&D, in-process control and release applications. Diagnostic applications are coming soon.
 
The Service includes:
  • Design and optimization of the PCR assay for the digital PCR analysis
  • Validation of the analytical procedure based on digital PCR methodology
  • Analysis of the samples
Potential applications are:
  • Quantification of the transgene copy number
  • Characterization of the reference material for the validation studies

 

How to Order

 

Every digital PCR project is different. Please ask our application specialists for your quote.

The following questions are relevant for appropriate pricing:

  • DNA/RNA isolation necessary? From what kind of organism and tissue?
  • Expected number of samples (single analysis, repeated analysis)?
  • Delivery of samples (amount, quality and vessel)?
  • Assay established?
  • Single or multiplex assay?
  • Number of replicas per sample?
  • Delivery form of data (raw data, analysed data, report)
  • Test samples available? Especially in case of larger projects, we prefer to perform initial testing in order to make a good bid.