Oscarliu117 (Talk | contribs) |
Oscarliu117 (Talk | contribs) |
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+ | <div id="list-item-5"> | ||
+ | |||
+ | </br> | ||
+ | <div class="row"> | ||
+ | <a class="btn col-md-12" data-toggle="collapse" href="#DNS" role="button" | ||
+ | aria-expanded="false" aria-controls="multiCollapseExample1"> | ||
+ | DNS reductive sugar measurement | ||
+ | <i class="fa fa-arrow-down fa-10" aria-hidden="true"></i> | ||
+ | </a> | ||
+ | </div> | ||
+ | |||
+ | <div class="collapse multi-collapse" id="DNS"> | ||
+ | <h3>DNS solution preparation:</h3></br> | ||
+ | <p class="pcontent">1. Disolve 2.5g of 3,5-Dinitrosalicylic acid (DNS) to 150ml | ||
+ | double distilled water.</br></br> | ||
+ | 2. Heat to solution to 45 degree Celsius and add 4g of NaOH. Stir the solution | ||
+ | until it is transparent.</br></br> | ||
+ | 3. Add 75g of potassium sodium tartrate and add water to 250 ml.</br></br> | ||
+ | 4. Keep the solution without light exposure. The solution can be used after 7 | ||
+ | days.</br></br></p> | ||
+ | |||
+ | <h3>Principle:</h3></br> | ||
+ | <p class="pcontent">Under base solution, DNS will turn to brown color while | ||
+ | reacting with reductive sugar in high temperature. In the specific temperature | ||
+ | range, the color will have linear relationship with the reductive sugar | ||
+ | concentration.</br></br></p> | ||
+ | |||
+ | <h3>Calibration:</h3></br> | ||
+ | <p class="pcontent">1. Prepare glucose or xylose water solution to the following | ||
+ | concentration: 0, 0.2, 0.4, 0.8, 1.0, 1.2, 1.6, 2 g/L</br></br> | ||
+ | 2. Take 200 ul of sample, add 200 ul of DNS solution.</br></br> | ||
+ | 3. Heat the sample at 100 degree Celsius for 10mins.</br></br> | ||
+ | 4. Cool down the sample with ice to room temperature.</br></br> | ||
+ | 5. Measure the optical density of the sample with 540nm light wavelength.</br></br> | ||
+ | 6. Draw the graph of sugar concentration with respect to optical density.</br></br></p> | ||
+ | |||
+ | <h3>Calibration results:</h3></br> | ||
+ | |||
+ | <img class="contentimg col-2" src="https://static.igem.org/mediawiki/2018/3/3e/T--NCKU_Tainan--home_xylose_new.jpg"> | ||
+ | <img class="contentimg col-2" src="https://static.igem.org/mediawiki/2018/a/aa/T--NCKU_Tainan--home_glucose.png"> | ||
+ | |||
+ | <h3>Measurement:</h3></br> | ||
+ | <p class="pcontent">1. Take 200 ul of sample, add 200 ul of DNS solution.</br></br> | ||
+ | 2. Heat the sample at 100 degree Celsius for 10mins.</br></br> | ||
+ | 3. Cool down the sample with ice to room temperature.</br></br> | ||
+ | 4. Measure the optical density of the sample with 540nm light wavelength.</br></br> | ||
+ | 5. Get the concentration of reductive sugar from the Calibration graph.</br></br></p> | ||
+ | |||
+ | |||
+ | |||
+ | |||
+ | </div> | ||
+ | |||
+ | |||
+ | </div> | ||
+ | |||
</div> | </div> |
Revision as of 02:34, 22 September 2018