Real-Time PCR (qPCR) Advanced (Advanced)- Hands on
- 3 days (2 days can be customized)
- Course fee
- This course is essential for Medical Technologist and Scientists who want to use this technology quantitatively or qualitatively in diagnostics or research.
- This course is an advanced course. No basics of Real-Time PCR theory or practical skills will be revisited during this course. We strongly recommend that delegates only register for this Advanced Real-Time PCR Course if they have comprehensive and detailed knowledge and experience in the Basics of Real-Time PCR.
- Delegates who do not have these skills are strongly advised to attend the Basics of Real-Time PCR Course to ensure that they are familiar with the detailed PCR theory and appropriate practical experience, as well as applications and advanced molecular biology calculations.
Topics to be covered with this training includes:
- Detection of PCR products in real-time.
- Methods in real-time PCR
- Basic terms used in real-time PCR
- Four elements for the Quantification of target amounts.
- Guidelines for the preparation of template, primers, and probes.
- Purification of nucleic acid templates.
- Handling and storing primers and probes
- Guidelines for successful whole transcriptome amplification
- The importance of RNA quality.
- Working with degraded RNA
- The amplification from low cell numbers
- Guidelines for successful real-time PCR
1- Primer design
2 -Reaction chemistry
3- SYBR Green-based detection in real-time PCR
4- Probe-based detection in real-time PCR
5- Real-time RT-PCR direct from cell lysates
6- Genomic DNA in real-time RT-PCR
- Guidelines for successful multiplex real-time PCR.
- Guidelines for designing primers and probes
- Guidelines for selecting appropriate reporter dyes and quenchers for the probes
- Choice of PCR buffer
- Guidelines for evaluating the performance of a multiplex assay
- How can I overcome challenges associated with PCR annealing?
- Guidelines for programming the real-time cycler
- Guidelines for analyzing data from a multiplex assay
- Guidelines for successful virus load quantitation
- Absolute vs. Relative Quantification.
- Experimental development and validation workflows.
- 6 critical factors to help you choose the right thermal cycler
Objectives and Expected Results
- This course is structured to expose the delegate to the advantages of using real-time PCR technology and chemistries whilst empowering the delegate with hands-on experience of real-time PCR applications.
- This course will enable scientists to understand the principles of designing an efficient real-time PCR assay, interpreting real-time PCR data, and troubleshooting real-time PCR assays.
Others Training Materials
- Workshop, Sheets, and case studies paper will be conducted and implemented during training.
- Documentary Videos will be presented.