Sanger Sequencing and Fragment Analysis by Capillary Electrophoresis (CE)

Overview

• Capillary electrophoresis (CE) enables accurate, unambiguous results for targeted genomic analysis.
• Two main methods: Sanger sequencing and fragment analysis.
• Both methods provide simple workflows, high accuracy, and fast turnaround.

Sanger Sequencing

• Gold-standard technology for accurate sequence determination with single-base resolution.
• Ideal for targeted sequencing and confirmation analysis.
• Supports various applications:
  • mRNA manufacturing QC
  • Gene editing confirmation (CRISPR, TALENs, etc.)
  • Cell line authentication
• InnoviGene Suite: A browser-based platform providing tools to automate and optimize data analysis.

Fragment Analysis

• Simple 4-step method for multiplexed detection of pathogens, variants, SNPs, or size analysis.
• Applications include:
  • Pathogen detection
  • Variant analysis
  • SNP sizing
• Software Tools: InnoviGene Suite for automated analysis.

Genetic Analysis Technologies

• Instruments include genetic analyzers, reagents, and software for both Sanger sequencing and fragment analysis.
• Software: Tools for viewing and interpreting CE results.
• Reagents: Available for purchase to support CE processes.

Applications in Clinical Research

• Sanger sequencing and fragment analysis are foundational in clinical research and labs.
• Applications:
  • mRNA manufacturing QC: Confirm sequence during therapeutic and vaccine production.
  • Gene Editing: Validation of genome engineering technologies like CRISPR.
  • Cell Line Authentication: STR genotyping for cell line verification.

Specific Applications and Case Studies

• DNA Methylation Analysis: Using Sanger sequencing for 5-meC detection; important in cancer research.
• Human Cell Line Authentication: STR profiling for cell line verification.
• Low-Level Somatic Variant Detection: Detecting mutant alleles in tumor tissues.
• Microbial Species Identification: Uses Sanger sequencing for species ID via rDNA sequences.
• Microsatellite Instability Analysis: TrueMark MSI Assay for tumor classification.
• Multiplex Ligation-Dependent Probe Amplification: For copy number variation analysis.
• Plasmid Sequencing: Analyzing inserts subcloned into plasmids.
• Repeat DNA Expansion: Analyzing challenging repeat-rich regions with SeqStudio.
• SARS-CoV-2 Identification: Multiplex qPCR solutions and fragment analysis for pathogen detection.
• SNP Genotyping: SNaPshot Multiplex System for analyzing genetic variants.

Human Identification

• CE and NGS applications in forensic DNA analysis.

Additional Resources

• Guides, webinars, application notes, and support for CE technologies.
• Training and publications available for further learning and application of CE technologies.

Choosing the Right Sequencing Method

• Important considerations provided for selecting appropriate sequencing methods based on application needs.

Educational and Support Resources

• CE training resources, learning centers, and Seq It Out video series.
• Double Helix Diaries podcast explores CE technology and applications.

Ordering and Support

• Thermo Fisher provides comprehensive support for CE product ordering and technical assistance.