Difference between revisions of "Team:Paris Bettencourt/Protocols"
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+ | <div class="textBox" id="VitaminATitration"> | ||
+ | <h3>Vitamin A Titration</h3> | ||
+ | <ul> | ||
+ | <li>Take 1g of food (with particle < 1mm of diameter) in bottle protected of the light, and add 15ml of hexane. Mix it.</li> | ||
+ | <li>Add again 15ml of hexane and shake during 5-10 minutes (do the same time, if you want compare two food sample).</li> | ||
+ | <li>In an other bottle protected of the light, do a filtration (coarse cellulose filter) of the previous solution.</li> | ||
+ | <li>Take the batter who don't pass the filter, and 15ml of hexane, shake 5 minutes, and filtered it like previously, to take all B-caroten as possible from the food sample.</li> | ||
+ | <li>Put in the fridge at -20°C to conserve it.</li> | ||
+ | <li>Obtain the result with a spectrophotometer with wavelenght of 450 nm (with a blank of hexane).</li> | ||
+ | </ul> | ||
+ | </div> | ||
+ | |||
+ | |||
+ | <div class="textBox" id="VitaminATitrationHPLC"> | ||
+ | <h3>Vitamin A Titration using HPLC</h3 | ||
+ | <ul> | ||
+ | <li>Suspend cells in 1 mL of sterile water.</li> | ||
+ | <li>Add 0.5 to 0.75 mm glass beads and vortex for 3 minutes.</li> | ||
+ | <li>Add 2.5 mL of 0.2% (wt/vol) pyrogallol dissolved in methanol and vortex for 3 minutes.</li> | ||
+ | <li>Add 1.25 mL of 60% (wt/vol) KOH and vortex for 10 seconds.</li> | ||
+ | <li>Incubate for 1 h at 75ºC, vortexing every 15 minutes.</li> | ||
+ | <li>Add appropriate amount of hexane.</li> | ||
+ | <li>Centrifuge tubes for 5 minutes at 2,800 rpm.</li> | ||
+ | <li>Pipette 1 mL of hexane into a cuvette. </li> | ||
+ | </ul> | ||
+ | </div> | ||
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<div class="innerBox cyan"> | <div class="innerBox cyan"> | ||
− | <a href="https://2015.igem.org/Team:Paris_Bettencourt/Protocols | + | <a href="https://2015.igem.org/Team:Paris_Bettencourt/Protocols#VitaminATitration"> |
− | <div class="ptext"><p>Titration | + | <div class="ptext"><p>Vitamin A Titration</p></div> |
− | <img src=" | + | <img src=""> |
</a> | </a> | ||
</div> | </div> | ||
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<div class="box"> | <div class="box"> | ||
<div class="innerBox cyan"> | <div class="innerBox cyan"> | ||
− | <a href="https://2015.igem.org/Team:Paris_Bettencourt/Protocols | + | <a href="https://2015.igem.org/Team:Paris_Bettencourt/Protocols#VitaminATitrationHPLC"> |
− | <div class="ptext"><p>Titration | + | <div class="ptext"><p>Vitamin A Titration using HPLC</p></div> |
− | <img src=" | + | <img src=""> |
</a> | </a> | ||
</div> | </div> |
Revision as of 09:32, 11 September 2015