20.109(S15):Diagnostic primer analysis (Day8)

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20.109(S15): Laboratory Fundamentals of Biological Engineering

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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

  1. Use any extra time to continue working on your bird microbiome analysis.

Homework

Due on M1D9

  1. Trees

Due on M2D3

  1. 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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