20.109(S17):Complete Western blot and induce DNA damage for survival and quantitative PCR assays (Day2)

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20.109(S17): Laboratory Fundamentals of Biological Engineering

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Schedule Spring 2017        Announcements        Assignments        Homework        Communication
       1. High-throughput ligand screening        2. Gene expression engineering        3. Biomaterials engineering              

Introduction

Protocols

Part 1: Induce DNA damage

Part 1a: Treat cells for survival assay

Part 1b: Treat cells for quantitative PCR assay

Part 2: Complete Western blot

Part 3: Design primers for quantitative PCR assay

Part 4: Discuss research paper

We will end today with a discussion of the Dietlein et al. research article. In their research, the authors completed a screen to examine 1319 cancer-associated genes from 67 cell lines to identify cancer-cell specific mutations that are associated with DNA-PKcs dependence or addiction.

Our paper discussion will assist you in writing a cohesive story that clearly reports the data and provides strong support for the conclusions made about the data. During the paper discussion, everyone is expected to participate - either by volunteering or by being called upon!

Introduction

Remember the key components of an introduction:

  • What is the big picture?
  • Is the importance of this research clear?
  • Are you provided with the information you need to understand the research?
  • Do the authors include a preview of the key results?

Results

Carefully examine the figures. First, read the captions and use the information to 'interpret' the data presented within the image. Second, read the text within the results section that describes the figure.

  • Do you agree with the conclusion(s) reached by the authors?
  • What controls are included and are they appropriate for the experiment performed?
  • Are you convinced that the data are accurate and/or representative?

Discussion

Consider the following components of a discussion:

  • Are the results summarized?
  • Did the authors 'tie' the data together into a cohesive and well-interpreted story?
  • Do the authors overreach when interpreting the data?
  • Are the data linked back to the big picture from the introduction?

Reagents

Navigation links

Next day: Assess cell survival and harvest RNA for quantitative PCR assay

Previous day: Confirm cell lines and practice tissue culture techniques
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