Explore the intricacies of RT-qPCR in gene expression analysis through this 45-minute webinar presented by Jennifer Sandoval, M.S., a Technical Scientist at Thermo Fisher Scientific. Delve into the critical aspects of the RT-qPCR workflow, including RNA isolation and reverse transcription, and their impact on accurate gene expression quantitation. Learn about reverse transcriptase attributes, such as RNase H activity, thermostability, processivity, sensitivity, and inhibitor tolerance. Discover strategies for working with challenging RNA samples, including low-input RNA, limited sample sources, and samples with high structural complexity. Address common issues like genomic DNA contamination and explore solutions for various sample types, including archived tissue, blood, and plant samples. Gain valuable insights to optimize your RT-qPCR experiments and push the boundaries of gene expression analysis.
Breaking Through RT-qPCR Boundaries in Gene Expression Analysis
Overview
Syllabus
Intro
Webinar Outline
Reverse Transcription Basics
Quantitative Reverse Transcription Polymerase Chain Reaction (RT-qPCR)
RT-qPCR Workflow
Reverse Transcription Components
Reverse Transcriptase Considerations
Reverse Transcriptase Attributes: RNase H Activity
Reverse Transcriptase Attributes: Thermostability
Reverse Transcriptase Attributes: Processivity
Reverse Transcriptase Attributes: Sensitivity
Reverse Transcriptase Attributes: Inhibitor Tolerance
How to Choose a Reverse Transcriptase?
Working with Challenging/Suboptimal RNA Samples
Detection of Low-input RNA and/or Low-abundance Genes
Working with Limited Sample Sources
Samples with High Structural Complexity
Working with (Archived) Tissue Samples: FFPE
Working with (Archived) Tissue Samples: Blood
Working with Plant Samples: Challenges
Working with Plant Samples: Inhibition
Difficult RNA Samples
Genomic DNA Contamination
gDNA Removal with DNases
gDNA Removal with ezDNase Enzyme
Conclusion
Thank You for Attending the Webinar
Taught by
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