Difference between revisions of "20.109(F16):Asses DNA repair variability with comet chip (Day4)"

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(→‎Part 1: Load cells and induce DNA damage)
(→‎Part 1: Load cells and induce DNA damage)
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For this experiment everyone will use H<sub>2</sub>O<sub>2</sub> to induce DNA damage in three Coriell cell lines to assess the DNA repair capacity variability in healthy individuals.  Use the following plate map to as you complete the below steps:
 
For this experiment everyone will use H<sub>2</sub>O<sub>2</sub> to induce DNA damage in three Coriell cell lines to assess the DNA repair capacity variability in healthy individuals.  Use the following plate map to as you complete the below steps:
  
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[[Image:Fa16 M2D4 repair assay plate map.png|thumb|center|400 px|'''Plate map for comet chip repair assay.''' The labels on the right refer to the Coriell cell lines and the labels on the bottom refer to the H<sub>2</sub>O<sub>2</sub> dose.  Grey wells will be empty for technical reasons.]]
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[[Image:Fa16 M2D4 repair assay plate map.png|thumb|center|500 px|'''Plate map for comet chip repair assay.''' The labels on the right refer to the Coriell cell lines and the labels on the bottom refer to the H<sub>2</sub>O<sub>2</sub> dose.  Grey wells will be empty for technical reasons.]]
  
 
You will load comet chips prepared by the teaching faculty using the procedure from [http://engineerbiology.org/wiki/20.109%28F16%29:Test_comet_chip_loading_variables_%28Day2%29#Part_2:_Load_comet_chip M1D2]; however, please note the following differences:
 
You will load comet chips prepared by the teaching faculty using the procedure from [http://engineerbiology.org/wiki/20.109%28F16%29:Test_comet_chip_loading_variables_%28Day2%29#Part_2:_Load_comet_chip M1D2]; however, please note the following differences:

Revision as of 17:18, 25 August 2016

20.109(F16): Laboratory Fundamentals of Biological Engineering

Engelward PNAS 2006.png

Schedule Fall 2016        Announcements        Assignments        Homework        Communication
       1. Measuring Genomic Instability        2. Manipulating Metabolism        3. Engineering Biomaterials              

Introduction

Protocols

Part 1: Load cells and induce DNA damage

For this experiment everyone will use H2O2 to induce DNA damage in three Coriell cell lines to assess the DNA repair capacity variability in healthy individuals. Use the following plate map to as you complete the below steps:

Plate map for comet chip repair assay. The labels on the right refer to the Coriell cell lines and the labels on the bottom refer to the H2O2 dose. Grey wells will be empty for technical reasons.

You will load comet chips prepared by the teaching faculty using the procedure from M1D2; however, please note the following differences:

  • You will not complete the lysis incubation immediately following the cell loading, rather this will be done after the DNA damage induction step.
  • For the cell number you will use 500 K/mL and for the loading time you will use 30 min.

Part 2: Use comet chip to assess repair capacity

Reagents

Navigation links

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