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Team:Imperial College London/Protocols Auxin
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<h2>Salkowski</h2> | <h2>Salkowski</h2> | ||
<hr style="color:#225323;"/> | <hr style="color:#225323;"/> | ||
| + | <p><b>To make reagent:<b></p> | ||
| + | <p>1. Dissolve 0.811g of anhydrous FeCl3 in 10ml H2O to obtain 0.5M solution</p> | ||
| + | <p>2. Add 1ml of FeCl3 0.5M solution to 50ml of 35% HClO4 </p> | ||
| + | <p>3. store at room temperature in absence of sunlight</p> | ||
| + | |||
| + | <p><b>To perform the assay:<b></p> | ||
| + | <p>1. Measure the OD of your auxin producing cells and control cells at 600 nm to account for any difference in growth.</p> | ||
| + | <p>2. Spin down each cell sample and take an aliquot of supernatant to filter with ...</p> | ||
| + | <p>3. Add salkowski reagent to the filtered supernatant in a ratio of 2:1</p> | ||
| + | <p>4. Leave in the dark for 25-30 mins and then measure OD at 530 nm. (You should be able to see a visible colour change to pink/red if auxin is present. </p> | ||
<h2>HPLC</h2> | <h2>HPLC</h2> | ||
<hr style="color:#225323;"/> | <hr style="color:#225323;"/> | ||
Revision as of 16:11, 15 September 2011
Protocols
This page lists all the protocols used in our project. We have classified them into five main categories as follow.
Auxin Xpress
Effect of Auxin on Plants
Split Root Experiment
Salkowski
To make reagent:
1. Dissolve 0.811g of anhydrous FeCl3 in 10ml H2O to obtain 0.5M solution
2. Add 1ml of FeCl3 0.5M solution to 50ml of 35% HClO4
3. store at room temperature in absence of sunlight
To perform the assay:
1. Measure the OD of your auxin producing cells and control cells at 600 nm to account for any difference in growth.
2. Spin down each cell sample and take an aliquot of supernatant to filter with ...
3. Add salkowski reagent to the filtered supernatant in a ratio of 2:1
4. Leave in the dark for 25-30 mins and then measure OD at 530 nm. (You should be able to see a visible colour change to pink/red if auxin is present.
