Difference between revisions of "20.109(F16):Module 1"
Noreen Lyell (Talk | contribs) (→Assignments) |
Noreen Lyell (Talk | contribs) (→Overview) |
||
| Line 14: | Line 14: | ||
In this module you will measure DNA repair using two assays: the CometChip and immuno-fluorescence. Your first task is to critically think through the development of the CometChip assay and determine which conditions provide the best results for loading mammalian cells into the device. To this end, you will consider variables that effect cell loading into the microwells of the CometChip. The data you collect will be used to determine the conditions for subsequent assays. | In this module you will measure DNA repair using two assays: the CometChip and immuno-fluorescence. Your first task is to critically think through the development of the CometChip assay and determine which conditions provide the best results for loading mammalian cells into the device. To this end, you will consider variables that effect cell loading into the microwells of the CometChip. The data you collect will be used to determine the conditions for subsequent assays. | ||
| − | Next, you will use the CometChip assay to measure DNA damage in response to chemical treatments and to assess repair capacity across different cell lines. Specifically, you will study the | + | Next, you will use the CometChip assay to measure DNA damage in response to chemical treatments and to assess repair capacity across different cell lines. Specifically, you will study the effect of oxidative stress in these experiments. Last, you will examine the effect of chemical treatment on the abundance of double strand breaks using an immuno-fluorescence approach. |
Revision as of 20:08, 31 August 2016
Contents
Module 1
Lecturer: Bevin Engelward
Instructors: Noreen Lyell, Leslie McClain and Maxine Jonas
TA: Molly Bird
Lab manager: Hsinhwa Lee
Overview
In this module you will measure DNA repair using two assays: the CometChip and immuno-fluorescence. Your first task is to critically think through the development of the CometChip assay and determine which conditions provide the best results for loading mammalian cells into the device. To this end, you will consider variables that effect cell loading into the microwells of the CometChip. The data you collect will be used to determine the conditions for subsequent assays.
Next, you will use the CometChip assay to measure DNA damage in response to chemical treatments and to assess repair capacity across different cell lines. Specifically, you will study the effect of oxidative stress in these experiments. Last, you will examine the effect of chemical treatment on the abundance of double strand breaks using an immuno-fluorescence approach.
Lab links: day by day
M1D1: Prepare CometChips
M1D2: Test CometChip loading variables
M1D3: Measure DNA damage with CometChip
M1D4: Assess DNA repair variability with CometChip
M1D5: Seed cells for immuno-fluorescence assay
M1D6: Complete immuno-fluorescence assay
M1D7: Data analysis
Assignments
Data summary
Mini-presentation
References
CometChip: A high-throughput 96-well platform for measuring DNA damage in microarrayed human cells. Journal of Visualized Experiments 92: 1-11. A video of the procedure can be found here.
CometChip: Single-cell microarray for high-throughput detection of DNA damage. Methods in Cell Biology 112: 247-268.
