Real-Time PCR hands-on training

Real-Time PCR has been used since the early 1990s and is a routine standard not only for a variety of diagnostic applications. It is often considered to be well-known and easily manageable, and little attention is sometimes given to the proper training of the lab personnel who use it in practice, whether for gene expression measurements by relative quantification methods or for example for quantification based on a standard curve in diagnosis or for forensic purposes.

In our course you will gain confidentiality in using your Real-Time PCR instrument, the ability to critically evaluate your results, and assure yourself of good working practices. No matter which instrument you have or for what purpose you intend to use or already use Real-Time PCR, the program of this course that takes place at your workplace is always tailored to your current situation and needs. The aim of the course is to provide an overview of technology, principles, concepts and applications of Real-Time PCR. Emphasis is placed typically primarily on experiment planning and correct interpretation of results, which are two areas where the most common mistakes occur.



Who should attend?

The course is suitable for anyone who is planning to use Real-Time PCR technology as well as for those who want to deepen their knowledge. Previous knowledge of Real-Time PCR technology is not required.

What will you learn?

  • How to design and run a Real-Time PCR experiment
  • How to evaluate results using various methods (relative quantification, standard curve quantification, allelic discrimination, high-resolution melting, etc.) including practical examples using appropriate data files
  • How to identify a problem in a particular experiment based on the interpretation of amplification plots and how to eliminate it
  • How to choose controls including endogenous ones
  • You will get information on the organization of the lab and the desirable lab practices
  • You will communicate with an experienced lecturer and discuss your questions in a team of your close colleagues
  • And many more



Course agenda:

We will review your results with you and analyze specific issues. We will discuss the possibilities your Real-Time PCR instrument offers with respect to your goals and applications. We can tailor the course program to your instrument, workplace and needs. You will tell us what you want to focus on.

Outline of the agenda (to be discussed):

  • Principles and use of Real-Time PCR
  • Basics of data analysis
    • amplification plots, how to obtain Cq values
    • melting curves analysis
  • Assay design and validation – design in silico
    • probes and unspecific dyes
    • repeats, SNPs, secondary structures
    • specificity
  • Optimization
    • standard and melting curves
    • amplification efficiency and how do I know it?
    • specificity, sensitivity
  • Quantification using standard curve
    • planning, conducting, analysing
  • Relative quantification of not only gene expression
    • quantification models
    • amplification efficiency correction
    • selection of reference genes
    • alternative strategies of normalization
    • data distribution, choosing the proper statistical test
    • p- value - interpretation of results
  • Practical recommendations
    • lab setup
    • instruments and software
    • dispensable and non-dispensable controls



Course Logistics:

  • Language: English
  • Venue: Your lab or workplace
  • Date: To be agreed
  • Number of participants: To be agreed, always only members of your team
  • Time range: To be agreed, typically 1-2 days
  • Price: Please request a quote
  • Note that during the course, laboratory experiments may be performed upon agreement.

There are no dates for this course

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