Project protocol - Contents

• Workflow and sampling
• Equipment and supplies
• Reagents and solutions
• Procedure: Alcohol consumption (drinking in the dark model)
• Data

Workflow and sampling

Step	Procedure	Equipment	Data collected*
1	Mice weighed on each of four consecutive test days	Scales	Body weight
2	Alcohol consumption measured (drinking in the dark model)	Drinking tubes	Fluid levels

*Supplementary data are available for this project. See Vanderlinden1 downloads.

Equipment and supplies

• Scales
• Drinking tubes (10 mL)

Reagents and solutions

• 20% ethanol (v/v in water)

Procedure: Alcohol consumption (drinking in the dark model)

1. Mice are weighed on each of four consecutive test days, 1 h prior to lights out.
2. Four hours after lights out, water bottles are replaced with drinking tubes containing 20% ethanol.
3. Two hours after introduction of the alcohol, the drinking tubes are removed and fluid levels recorded.
4. On days 1-3, drinking tubes are replaced with water bottles.
5. On day 4, alcohol intake is determined for a second time (2 h later than previous measurement), and then tubes are then replaced with water bottles.

Data collected by investigator

• body weight on day 1
• ethanol consumption on days 2, 3, 4 (drinking in the dark)
• first principal component on log-transformed ethanol consumption data from days 2, 3, 4 (drinking in the dark)