Difference between revisions of "20.109(F16):Test comet chip loading variables (Day2)"
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#*Now that you have an idea as to the number of cells that are ideal for loading, consider the conditions you will use in your experiment. | #*Now that you have an idea as to the number of cells that are ideal for loading, consider the conditions you will use in your experiment. | ||
#**If you and your partner are in agreement on how many cells are ideal, you can load the ideal number of cells for Condition A and then either too few or too many for Condition B. Or you can load slightly more than the ideal number of cells for Condition A and slightly less for Condition B. | #**If you and your partner are in agreement on how many cells are ideal, you can load the ideal number of cells for Condition A and then either too few or too many for Condition B. Or you can load slightly more than the ideal number of cells for Condition A and slightly less for Condition B. | ||
− | #**If you and your partner are not in agreement, you can each load the number of cells you determine to be ideal as the | + | #**If you and your partner are not in agreement, you can each load the number of cells you determine to be ideal as the Condition A and Condition B. |
'''Experiment 2: Loading time''' | '''Experiment 2: Loading time''' |
Revision as of 19:23, 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 or conditions 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. In the comet chip that you prepared during the previous laboratory class there is space for you to complete two experiments, each with two conditions.In the comet chip that you prepared during the previous laboratory class there is space for you to complete two experiments, each with two conditions. Your experiments will address the questions above and the conditions will provide data that will, hopefully, answer your research questions.
Experiment 1: Cell number
- 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. Also, use the data you collected during the Orientation exercise to determine how many cells can fit into a single microwell based on the dimensions provided above.
- When you know how many cells you want to load into each microwell, move on to the next question.
- How many cells should be added to each well such that the desired number of cells are loaded into the microwells?
- Consider the likelihood that every cell you add to the well will fall into a microwell. Perhaps calculate the surface area of the bottom of a well and compare this to the size of the cells as you consider this question.
- Now that you have an idea as to the number of cells that are ideal for loading, consider the conditions you will use in your experiment.
- If you and your partner are in agreement on how many cells are ideal, you can load the ideal number of cells for Condition A and then either too few or too many for Condition B. Or you can load slightly more than the ideal number of cells for Condition A and slightly less for Condition B.
- If you and your partner are not in agreement, you can each load the number of cells you determine to be ideal as the Condition A and Condition B.
Experiment 2: Loading time
Part 2: Load comet chip
Reagents
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