IGEM:IMPERIAL/2007/Projects/Experimental Design/Improve Methodology/Protocol2

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 then be left overnight and measured again in the morning to determine the rate of evaporation. This rate will then be compared to the control which is just water with no oil.

Reagents

• Distilled Water x 1200ul
• Parafin Oil x 270ul
• Mineral Oil x 270ul

Equipment

• Pipettes P5,P20,P200
• 96 Well Plate (black)
• Fluorometer
• Incubator 37°C

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. Place the plate in the incubator at 37^oC

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 total 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 total sample volume and adjust accordingly. Remember that in a well samples can get stuck to the rim of the well.

Well	Water Added(ul)	Paraffin Oil	Total Volume (ul)
E5	60ul	10ul	70ul
F5	60ul	10ul	70ul
G5	60ul	10ul	70ul
E6	60ul	20ul	80ul
F6	60ul	20ul	80ul
G6	60ul	20ul	80ul
E7	60ul	90ul	150ul
F7	60ul	90ul	150ul
G7	60ul	90ul	150ul
B5	60ul	10ul	70ul
C5	60ul	10ul	70ul
D5	60ul	10ul	70ul
B6	60ul	20ul	80ul
C6	60ul	20ul	80ul
D6	60ul	20ul	80ul
B7	60ul	90ul	150ul
C7	60ul	90ul	150ul
D7	60ul	90ul	150ul