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:
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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:
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*You will not complete the lysis incubation immediately following the cell loading, rather this will be done after the DNA damage induction step.
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*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===
 
===Part 2: Use comet chip to assess repair capacity===

Revision as of 16:56, 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:


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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