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Seed germination "roll-ups" protocol for root hair measurement

Step 1: Sterilize seeds with 0.5% bleach for 5 minutes, if desired. Rinse 3 times with deionized water. If the seed is not treated, apply fungicide for 1 minute.

Step 2: Lay brown germination paper (Number 78, Anchor Paper Company, St. Paul, MN) on flat surface. Spray paper with 0.5 mM calcium sulfate solution until evenly damp. 

Step 3: Place 5 seeds in a row evenly spaced 2 cm from the top of the paper. Roll up paper with seeds inside. 

Step 4: Place roll(s) upright in a 2L beaker or container. Add 0.5 L 0.5 mM calcium sulfate solution in bottom of container.

Step 5: Place in germination chamber at 28 C for 4 days, or until seedlings emerge from top of roll. To prevent growth of fungus, take rolls out immediately after plants have emerged.

Step 6: Continue to grow seedlings in roll-ups in light chamber at 30 C for up to 10 days.