20.109(S15):Diagnostic primer analysis (Day8)
Introduction
Real-time reverse transcription-polymerase chain reaction (RRT-PCR) allows researchers to monitor the results of PCR as amplification is occurring (this technique is also referred to as quantitative or real-time PCR). During RRT-PCR data are collected throughout the amplification process using a fluorescent dye. The fluorescent dye is highly specific for double-stranded DNA and when bound to DNA molecules the fluorescence intensity increases proportionate to the increase in double-stranded product. In contrast, the data for traditional PCR are simply observed as a band on a gel (remember back to M1D5).
An animation of this process is linked here. http://www.sigmaaldrich.com/life-science/molecular-biology/pcr/quantitative-pcr/sybr-green-based-qpcr/syber-green-animation.html
Protocols
Part 1: AIV screening analysis
Part 2: Bird microbiome analysis
- Use any extra time to continue working on your bird microbiome analysis.
Homework
Due on M1D9
- Trees
Due on M2D3
- You will report your findings for the AIV screening portion of this module as a Primer Design Memo. Review the guidelines for this assignment and feel free to get an early start!
Reagent List
- Your brains!
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Previous Day: Phylogenetic analysis