Difference between revisions of "20.109(F16):Test comet chip loading variables (Day2)"
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'''Experiment 1: Cell number'''<br> | '''Experiment 1: Cell number'''<br> | ||
| − | This first experiment has two questions that should be considered as you discuss the variables you will test with your partner. Before moving on to these questions, it is important to differentiate between the terms 'well' and 'microwell' for | + | This first experiment has two questions that should be considered as you discuss the variables you will test with your partner. Before moving on to these questions, it is important to differentiate between the terms 'well' and 'microwell' for your experiments. Bottomless 96-well plates are placed on top of the agarose comet chip to create the wells for the comet chip assay (panel A). This enables researchers to control which cells are exposed to which treatment. The microwells were stamped into the agarose when you made your comet chip. Within each well is XX microwells, which are X x X &u;m. |
#How many cells should be loaded into each microwell? | #How many cells should be loaded into each microwell? | ||
Revision as of 18:11, 30 June 2016
Contents
Introduction
experimental design...
Protocols
Part 1: Design experiment to optimize comet chip loading
You will design two experiments that test variables associated with loading cells into the microwells of your comet chip. In the first experiment you will consider the role of cell number. Specifically, how many cells should be added to each well to ensure the majority of the microwells are loaded? In addition, how many cells should be loaded into each microwell? The second experiment will interrogate the length of time needed for loading.
Experiment 1: Cell number
This first experiment has two questions that should be considered as you discuss the variables you will test with your partner. Before moving on to these questions, it is important to differentiate between the terms 'well' and 'microwell' for your experiments. Bottomless 96-well plates are placed on top of the agarose comet chip to create the wells for the comet chip assay (panel A). This enables researchers to control which cells are exposed to which treatment. The microwells were stamped into the agarose when you made your comet chip. Within each well is XX microwells, which are X x X &u;m.
- How many cells should be loaded into each microwell?
- Consider the amount of DNA that is carried by a single mammalian cell and the detection limit provided by the SYBR gold DNA stain that will be used in your experiments.
- How many cells should be added to each well such that the desired number of cells are loaded into the microwells?
Experiment 2: Loading time
Part 2: Load comet chip
Reagents
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