IGEM:IMPERIAL/2007/Projects/Experimental Design/Improve Methodology/Protocol2: Difference between revisions

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===Testing Evaporation when using Oil===
===Testing Evaporation when using Oil===
We have decided to test how we can prevent evaporation using oragnic oils. Two types of oil will be tested, parafin oil and mineral oil, at varying volumes to see which is a suitable volume of oil to use.
We have decided to test how we can prevent evaporation using oragnic oils. Two types of oil will be tested, parafin oil and mineral oil, at varying volumes to see which is a suitable volume of oil to use.

Revision as of 06:42, 12 September 2007

Testing Evaporation when using Oil

We have decided to test how we can prevent evaporation using oragnic oils. Two types of oil will be tested, parafin oil and mineral oil, at varying volumes to see which is a suitable volume of oil to use. We will be testing 60ul of water with varying volumes of mineral oil and parafin oil. The samples will be left overnight and measured again in the morning to determine the rate of evaporation. This rate will be compared to the control which is just water on its own

Reagents

  • Distilled Water
  • Parafin Oil
  • Mineral Oil

Equipment

  • Pipettes P5,P20,P200
  • Plate
  • Fluorometer

Protocol

Day 1
Following the following schemetic first pipette 60ul of water into each well. Then pipette in the required volume and type of oil Note that oil can be tricky to pipette because of the capillary action of pipettes, be aware of this.
Day 2
To measure the evaporation we use a gilson pipette. To do this adjust the volume to a suitable volume and remove the sample until all is removed, note down the volume removed and minus this from the original volume. When we carried this out we started at 20ul and then adjusted down to 5ul when 20ul was no longer appropriate. It is best to judge the measurement based on the sample and adjust accordingly. Remember that in a well samples can get stuck to the rim of the well and so it is important to remove this as well.

Well Test Construct In vitro chassis Vol of in
vitro chassis (ul)
e5 60ul 10ul of Parafin oil 70ul
f5 60ul 10ul of Parafin oil 70ul
g5 60ul 10ul of Parafin oil 70ul
e6 60ul 20ul of Parafin oil 80ul
f6 60ul 20ul of Parafin oil 80ul
g6 60ul 20ul of Parafin oil 80ul
e7 60ul 90ul of Parafin oil 150ul
f7 60ul 90ul of Parafin oil 150ul
g7 60ul 90ul of Parafin oil 150ul
b5 60ul 10ul of Parafin oil 70ul
c5 60ul 10ul of Parafin oil 70ul
d5 60ul 10ul of Parafin oil 70ul
b6 60ul 20ul of Parafin oil 80ul
c6 60ul 20ul of Parafin oil 80ul
d6 60ul 20ul of Parafin oil 80ul
b7 60ul 90ul of Parafin oil 150ul
c7 60ul 90ul of Parafin oil 150ul
d7 60ul 90ul of Parafin oil 150ul