20.109(F16):Test comet chip loading variables (Day2)

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 our experiments.

1. 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.

1. How many cells should be added into the well such that the desired number of cells are loaded into the microwell?

Experiment 2: Loading time

Part 2: Load comet chip

Reagents

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