The Essential DNA Sequencing course is part of the Essential Short Course Series™ and is held over a period of three and a half days.
Validation of molecular assays is an essential component of quality assurance. At the molecular level sequencing generates absolute proof of DNA alterations. For this reason, DNA sequencing remains an essential and powerful tool for the molecular biologist. This course will highlight the power of DNA sequence information. Delegates will be trained according to an automated sequence protocol. This hands-on course will empower the delegate with all the necessary theoretical and practical knowledge to use the tool of DNA sequencing to its fullest potential. The use of sequencing services will be highlighted, enabling analysts to generate sequencing data without owning a sequencer.
The Essential DNA Sequencing course is an advanced course. No basic PCR theory or practical skills will be revisited during course, and we strongly recommend that delegates only register for this course if they have comprehensive and detailed knowledge and experience in PCR.
Delegates who do not have these skills are advised to attend the Essential PCR course to ensure that they are familiar with the detailed PCR theory and have the appropriate practical experience, as well as applications and advanced molecular biology calculations.
Course Programme
Day 1
Introduction to DNA sequencing theory and related topics
- Brief overview of DNA and gene structure
- Principles of automated DNA sequencing
- Step-for-step discussion of the automated DNA sequencing protocol
Discussions of Practical Sessions
- Planning and execution of a sequencing experiment
Practical Session I Hands-on experience
- PCR the fragment that will subsequently be sequenced
- Verify quality of amplification product for sequencing
Practical Session II Hands-on experience
- Perform automated sequencing reaction
Day 2
Analysis of sequence data
- Detailed analysis of sequence data generated in practical session
- Multiple sequence analysis tools will be demonstrated
Practical Session III Hands-on experience
- Perform PCR purification of PCR product
- Assess purified PCR product
Practical Session IV Hands-on experience
- Perform automated sequencing
Fluorescent labelling
- Dye labelled components
- Terminator chemistry
- Different platforms available for sequencing
Automated Sequencing
- Cycle sequencing
- Components of the reaction
- Preparation of sequenced product
- Electrophoresis
- Filter sets
- Matrix files
Day 3
Analysis of sequence data
- Detailed analysis of sequence data generated in practical session
Sequence Data A-Z
- Storing sequence data
- Data formats
- Editing programmes
- Alignments
- Databases
- DNA / genomic
- Protein sequence and structure
- Annotation of databases
- BLAST services
- Step-for-step demonstration of selected databases and programmed
Practical Session V Hands-on experience
- Perform automated sequencing
Discussions
- Manual sequencing
- Ethical principles related to sequence information
Day 4
Analysis of sequence data
- Detailed analysis of sequence data generated in practical session
Troubleshoot of automated sequencing results
- Scope of the problem and where to start troubleshooting
- More than 10 examples are discussed
Discussion
- Quality assurance and sequencing
- Validation of molecular assays – the role of sequence data
The sequence of this programme is preliminary and may change.