Oscarliu117 (Talk | contribs) |
Oscarliu117 (Talk | contribs) |
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</br> | </br> | ||
<h3>DNS solution preparation:</h3></br> | <h3>DNS solution preparation:</h3></br> | ||
− | <p class="pcontent"> | + | <p class="pcontent"> |
− | + | <ol> | |
− | + | <li>Disolve 2.5g of 3,5-Dinitrosalicylic acid (DNS) to 150ml | |
− | + | double distilled water.</li></br> | |
− | + | <li>Heat to solution to 45 degree Celsius and add 4g of NaOH. Stir the solution | |
− | + | until it is transparent.</li></br> | |
− | + | <li>Add 75g of potassium sodium tartrate and add water to 250 ml.</li></br> | |
+ | <li>Keep the solution without light exposure. The solution can be used after 7 | ||
+ | days.</li></br> | ||
+ | </ol> | ||
+ | </p> | ||
<h3>Principle:</h3></br> | <h3>Principle:</h3></br> | ||
− | <p class="pcontent">Under base solution, DNS will turn to brown color while | + | <p class="pcontent"> |
+ | Under base solution, DNS will turn to brown color while | ||
reacting with reductive sugar in high temperature. In the specific temperature | reacting with reductive sugar in high temperature. In the specific temperature | ||
range, the color will have linear relationship with the reductive sugar | range, the color will have linear relationship with the reductive sugar | ||
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<h3>Calibration:</h3></br> | <h3>Calibration:</h3></br> | ||
− | <p class="pcontent"> | + | <p class="pcontent"> |
− | + | ||
− | + | <ol> | |
− | + | <li>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.</li></br> | |
− | + | <li>Take 200 ul of sample, add 200 ul of DNS solution.</li></br> | |
− | + | <li>Heat the sample at 100 degree Celsius for 10mins.</li></br> | |
+ | <li>Cool down the sample with ice to room temperature.</li></br> | ||
+ | <li>Measure the optical density of the sample with 540nm light wavelength.</li></br> | ||
+ | <li>Draw the graph of sugar concentration with respect to optical density.</li></br> | ||
+ | </ol> | ||
+ | </p> | ||
<h3>Calibration results:</h3></br> | <h3>Calibration results:</h3></br> | ||
− | + | ||
− | + | ||
− | + | <img class="contentimg" src="https://static.igem.org/mediawiki/2018/f/fa/T--NCKU_Tainan--home_42846829_332575510640801_8298239152197992448_n.png"></br></br> | |
− | + | ||
− | + | ||
− | + | ||
<h3>Measurement:</h3></br> | <h3>Measurement:</h3></br> | ||
− | <p class="pcontent"> | + | <p class="pcontent"> |
− | + | <ol> | |
− | + | <li>Take 200 ul of sample, add 200 ul of DNS solution.</li></br> | |
− | + | <li>Heat the sample at 100 degree Celsius for 10mins.</li></br> | |
− | + | <li>Cool down the sample with ice to room temperature.</li></br> | |
+ | <li>Measure the optical density of the sample with 540nm light wavelength.</li></br> | ||
+ | <li>Get the concentration of reductive sugar from the Calibration graph.</li></br> | ||
+ | </ol> | ||
</p> | </p> | ||
</div> | </div> |
Revision as of 12:25, 29 September 2018