Oscarliu117 (Talk | contribs) |
Oscarliu117 (Talk | contribs) |
||
Line 4: | Line 4: | ||
<head> | <head> | ||
− | <link rel="stylesheet" href="https://2018.igem.org/Template:NCKU_Tainan/css/ | + | <link rel="stylesheet" href="https://2018.igem.org/Template:NCKU_Tainan/css/HP_Integrated_Human_Practices_style?action=raw&ctype=text/css"> |
</head> | </head> | ||
− | <body> | + | <body data-spy="scroll" data-target=".navbar-example"> |
+ | |||
+ | <header> | ||
+ | <div class="carousel-inner" role="listbox"> | ||
+ | <div class="carousel-item active" style="background-image: url('http://placehold.it/1900x1080')"> | ||
+ | </div> | ||
+ | </div> | ||
+ | </header> | ||
− | |||
<div class="container content"> | <div class="container content"> | ||
+ | <h1 class="head">Integrated Human Practices</h1> | ||
<div class="navbar-example"> | <div class="navbar-example"> | ||
<div class="row"> | <div class="row"> | ||
− | + | <div class="col-2 side"> | |
− | + | <div id="sidelist" class="list-Fgroup"> | |
− | + | <a class="list-group-item list-group-item-action" href="#list-item-1">Professor</a> | |
− | + | <a class="list-group-item list-group-item-action" href="#list-item-2">Meet up</a> | |
− | + | <a class="list-group-item list-group-item-action" href="#list-item-3">Enterprise Visit</a> | |
− | + | <a class="list-group-item list-group-item-action" href="#"><i class="fa fa-arrow-up fa-1x" | |
− | + | aria-hidden="true"></i></a> | |
− | + | ||
− | + | ||
− | + | ||
− | + | ||
</div> | </div> | ||
− | + | </div> | |
− | <div class="col- | + | <div class="col-10"> |
<div data-spy="scroll" data-target="#sidelist" data-offset="0" class="scrollspy-example"> | <div data-spy="scroll" data-target="#sidelist" data-offset="0" class="scrollspy-example"> | ||
− | |||
− | + | <div id="list-item-1"> | |
− | + | </br> | |
− | + | <div class="row"> | |
− | + | <a class="btn col-md-12" data-toggle="collapse" href="#PCR" role="button" aria-expanded="false" | |
− | + | aria-controls="multiCollapseExample1"> | |
− | + | PCR | |
− | + | <i class="fa fa-arrow-down fa-10" aria-hidden="true"></i> | |
− | + | </a> | |
− | + | </div> | |
− | + | <div class="collapse multi-collapse" id="PCR"> | |
− | + | <p class="pcontent"></br></br>1. Gently mix the following reaction by pipetting and | |
− | + | centrifuge briefly.</p> | |
− | + | <table class="card card-body"> | |
− | + | <tr> | |
− | + | <th></th> | |
− | + | <th>20 μl system</th> | |
− | + | <th>50 μl system</th> | |
− | + | </tr> | |
− | + | <tr> | |
− | + | <td>Template</td> | |
− | + | <td>12~20 ng</td> | |
− | + | <td>30~50 ng</td> | |
− | + | </tr> | |
− | + | <tr> | |
− | + | <td>Forward primer</td> | |
− | + | <td>1.0 μl</td> | |
− | + | <td>2.5 μl</td> | |
− | + | </tr> | |
− | + | <tr> | |
− | + | <td>Reverse primer</td> | |
− | + | <td>1.0 μl</td> | |
− | + | <td>2.5 μl</td> | |
− | + | </tr> | |
− | + | <tr> | |
− | + | <td>dNTP</td> | |
− | + | <td>1.6 μl</td> | |
− | + | <td>4.0 μl</td> | |
− | + | </tr> | |
− | + | <tr> | |
− | + | <td>10x Buffer</td> | |
− | + | <td>2.0 μl</td> | |
− | + | <td>5.0 μl</td> | |
− | + | </tr> | |
− | + | </table> | |
− | + | <p class="pcontent"></br>Program of KOD DNA polymerase</p> | |
− | + | <table class="card card-body"> | |
− | + | <tr> | |
− | + | <th>Temperature</th> | |
− | + | <th>Time</th> | |
− | + | <th></th> | |
− | + | </tr> | |
− | + | <tr> | |
− | + | <td>94 ℃</td> | |
− | + | <td>3 min.</td> | |
− | + | <td rowspan="3">25~30 cycles</td> | |
− | + | </tr> | |
− | + | <tr> | |
− | + | <td>94 ℃</br>(Denaturation)</td> | |
− | + | <td>40 sec</td> | |
− | + | </tr> | |
− | + | <tr> | |
− | + | <td>57.5 ℃</br>(Annealing)</td> | |
− | + | <td>30 sec</td> | |
− | + | </tr> | |
− | + | <tr> | |
− | + | <td>72 ℃</br>(Extension)</td> | |
− | + | <td>Depend on sequence size</br>(2 kbp/min. for Taq)</td> | |
− | + | <td></td> | |
− | + | </tr> | |
− | + | <tr> | |
− | + | <td>72 ℃</td> | |
− | + | <td>5 min.</td> | |
− | + | <td></td> | |
− | + | </tr> | |
− | + | <tr> | |
− | + | <td>4 ℃</td> | |
− | + | <td>∞</td> | |
− | + | <td></td> | |
− | + | </tr> | |
− | + | </table> | |
− | + | <p class="pcontent"></br>2. Confirm the size of the digested product by gel | |
− | + | electrophoresis.</br></br> | |
− | + | 3. Gel purification of the target size.</br></br></p> | |
− | + | </div> | |
+ | </div> | ||
+ | |||
+ | <div id="list-item-2"> | ||
+ | |||
+ | </br> | ||
+ | <div class="row"> | ||
+ | <a class="btn col-md-12" data-toggle="collapse" href="#Plasmid_Construction" role="button" | ||
+ | aria-expanded="false" aria-controls="multiCollapseExample1"> | ||
+ | Plasmid Construction | ||
+ | <i class="fa fa-arrow-down fa-10" aria-hidden="true"></i> | ||
+ | </a> | ||
</div> | </div> | ||
− | <div id=" | + | <div class="collapse multi-collapse" id="Plasmid_Construction"> |
+ | </br></br> | ||
+ | <p class="pcontent">1. Digestion (vector)</p> | ||
+ | <table class="card card-body"> | ||
+ | <tr> | ||
+ | <th>Plasmid</th> | ||
+ | <th>200 ng</th> | ||
+ | <th>1000 ng</th> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td>EcoRI / SpeI</td> | ||
+ | <td>0.2 μl</td> | ||
+ | <td>1 μl</td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td>XbaI / PstI</td> | ||
+ | <td>0.2 μl</td> | ||
+ | <td>1 μl</td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td>CutSmart Buffer</td> | ||
+ | <td>2 μl</td> | ||
+ | <td>5 μl</td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td>ddH2O</td> | ||
+ | <td>Up to 20 μl</td> | ||
+ | <td>Up to 50 μl</td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td>Digestion at 37℃ for 2.5hr.</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | |||
+ | </table> | ||
</br> | </br> | ||
− | < | + | <p class="pcontent">2. Digestion (insert)</p> |
− | + | <table class="card card-body"> | |
− | + | <tr> | |
− | Plasmid | + | <th>Plasmid</th> |
− | < | + | <th>200 ng</th> |
− | + | <th>1000 ng</th> | |
− | + | </tr> | |
− | + | <tr> | |
− | < | + | <td>EcoRI / XbaI</td> |
− | + | <td>0.2 μl</td> | |
− | + | <td>1 μl</td> | |
− | < | + | </tr> |
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | <tr> | |
− | + | <td>SpeI / PstI</td> | |
− | + | <td>0.2 μl</td> | |
− | + | <td>1 μl</td> | |
− | + | </tr> | |
− | + | <tr> | |
− | + | <td>CutSmart Buffer</td> | |
− | + | <td>2 μl</td> | |
− | + | <td>5 μl</td> | |
− | + | </tr> | |
− | + | <tr> | |
− | + | <td>ddH2O</td> | |
− | + | <td>Up to 20 μl</td> | |
− | + | <td>Up to 50 μl</td> | |
− | + | </tr> | |
− | + | <tr> | |
− | + | <td>Digestion at 37℃ for 2.5hr.</td> | |
− | + | <td></td> | |
− | + | <td></td> | |
− | + | </tr> | |
− | + | </table> | |
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | </ | + | </br> |
+ | <p class="pcontent">3.Confirm the size of the digested product by gel | ||
+ | electrophoresis.</br></br> | ||
+ | 4. Gel purification of the target size.</br></br> | ||
+ | 5. Ligation</br> | ||
− | |||
− | |||
<table class="card card-body"> | <table class="card card-body"> | ||
+ | |||
<tr> | <tr> | ||
− | < | + | <td>Vector (2 kbp)</td> |
− | < | + | <td rowspan="2">molar ratio = 1:3</br>(can up to 1:10 depending on the |
− | + | DNA sizes)</td> | |
</tr> | </tr> | ||
<tr> | <tr> | ||
− | <td> | + | <td>Insert (1.5 kbp)</td> |
− | + | ||
− | + | ||
</tr> | </tr> | ||
− | + | <tr> | |
− | <td> | + | <td>Quick Ligase Reaction Buffer (2X)*</td> |
− | + | <td>10 μl</td> | |
− | <td> | + | |
</tr> | </tr> | ||
<tr> | <tr> | ||
− | <td> | + | <td>Quick Ligase</td> |
− | + | <td>1 μl</td> | |
− | + | ||
</tr> | </tr> | ||
− | + | <tr> | |
<td>ddH2O</td> | <td>ddH2O</td> | ||
− | + | <td>Up to 20 μl</td> | |
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
</tr> | </tr> | ||
Line 228: | Line 265: | ||
</br> | </br> | ||
− | <p class="pcontent"> | + | <p class="pcontent">6. Transform the product by heat shock.</br> |
− | + | ||
− | + | ||
− | + | ||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | + | </div> | |
− | + | </div> | |
− | + | ||
− | |||
− | |||
− | |||
− | |||
− | + | <div id="list-item-3"> | |
− | + | </br> | |
− | + | <div class="row"> | |
− | + | <a class="btn col-md-12" data-toggle="collapse" href="#Extraction" role="button" | |
+ | aria-expanded="false" aria-controls="multiCollapseExample1"> | ||
+ | PCR Clean-Up & Gel Extraction | ||
+ | <i class="fa fa-arrow-down fa-10" aria-hidden="true"></i> | ||
+ | </a> | ||
+ | </div> | ||
− | + | <div class="collapse multi-collapse" id="Extraction"> | |
− | + | ||
− | + | ||
− | + | ||
− | + | </br></br> | |
+ | <h3>Gel Dissociation</h3></br> | ||
+ | <p class="pcontent">1. Gel Extraction</br></br> | ||
+ | a. Excised the DNA fragment from the agarose gel.</br></br> | ||
+ | b. Transferred up to 300 mg of the gel slice to a 1.5 ml microcentrifuge tube.</br></br> | ||
+ | c. Added 500 μl of the Gel/PCR Bufffer to the sample and mixed by vortex. | ||
+ | Incubate | ||
+ | at 55~60℃ for 10 minutes (or until the gel slice has completely dissolved).</br></br> | ||
+ | d. During the incubation, mixed by vortexing the tube every 2~3 minutes.</br></br> | ||
+ | e. Cooled the dissolved sample mixture to the room temperature.</br></br> | ||
+ | </p> | ||
+ | <h3>DNA Binding</h3></br> | ||
+ | <p class="pcontent">2. Placed a PG Column in a Collection Tube. Apply the | ||
+ | supernatant to the PG | ||
+ | Column | ||
+ | by decanting or pipetting.</br></br> | ||
+ | 3. Centrifuged at 16,000 xg for 30 seconds.</br></br> | ||
+ | 4. Discarded the flow-through and place the PG Column back into the same | ||
+ | collection | ||
+ | tube.</br></br></p> | ||
+ | <h3>Wash</h3></br> | ||
+ | <p class="pcontent">6. Added 400 μl of the Buffer W1 into the PG Column.</br></br> | ||
+ | 7. Centrifuged at 16,000 xg for 30 seconds.</br></br> | ||
+ | 8. Discarded the flow-through and place the PG Column back into the same | ||
+ | collection | ||
+ | tube.</br></br> | ||
+ | 9. Added 600 μl of the Buffer W2 (ethanol added) into the PG Column.</br></br> | ||
+ | 10. Centrifuged at 16,000 xg for 30 seconds.</br></br> | ||
+ | 11. Discarded the flow-through and place the PG Column back into the same | ||
+ | collection tube.</br></br> | ||
+ | 12. Centrifuged at 16,000 xg again for 2 minutes to remove the residual Buffer | ||
+ | W2.</br></br></p> | ||
+ | <h3>Elution</h3></br> | ||
+ | <p class="pcontent">13. To elute the DNA, placed the PG Column in a clean 1.5 ml | ||
+ | microcentrifuge | ||
+ | tube.</br></br> | ||
+ | 14. Added 50 μl of the H2O (pH is between 7.0 and 8.5) to the center of each PG | ||
+ | Column, let it stand for at least 2 minutes, and centrifuge at 16,000 xg for 2 | ||
+ | min.</br></br></p> | ||
+ | </p> | ||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
</div> | </div> | ||
+ | </div> | ||
+ | </br> | ||
+ | <div class="row"> | ||
+ | <a class="btn col-md-12" data-toggle="collapse" href="#Preparation" role="button" | ||
+ | aria-expanded="false" aria-controls="multiCollapseExample1"> | ||
+ | Competent Cell Preparation | ||
+ | <i class="fa fa-arrow-down fa-10" aria-hidden="true"></i> | ||
+ | </a> | ||
+ | </div> | ||
− | + | <div class="collapse multi-collapse" id="Preparation"> | |
− | </br> | + | <p class="pcontent"></br></br>・ For E. coli DH5α,BL21(DE3) and W3100(DE3) competent |
− | < | + | cell</br></br> |
− | + | 1. Streak out wild type E. coli on a plate (LB plate without antibiotics) overnight | |
− | + | and pick one colony into 3 ml of media (LB) and grow overnight.</br></br> | |
− | + | 2. Transfer 0.4 ml of starter culture into 40 ml of fresh LB and grow culture at 37 | |
− | + | ℃.</br></br> | |
− | + | 3. When the OD600 nm up to 0.35, put the cells on ice immediately.</br></br> | |
− | </ | + | 4. Spin the cells at 4℃for 10 minutes at 4000 rpm.</br></br> |
+ | 5. Suspend the pellet on ice carefully with 16 ml chilly Transformation Buffer | ||
+ | 1(TFB1)</br></br> | ||
+ | 6. Leave nicely suspended bugs on ice for 10 minutes.</br></br> | ||
+ | 7. Spin the cells at 4℃ for 10 min. at 4000 rpm.</br></br> | ||
+ | 8. Suspend the pellet on ice with 1.6 ml of Transformation Buffer 2 (TFB2).</br></br> | ||
+ | 9. Leave on immediately on ice for 30 minutes.</br></br> | ||
+ | 10. Aliquot 100 μl into 1.5 ml centrifuge tubes and snap freeze immediately with | ||
+ | liquid nitrogen.</br></br> | ||
+ | 11. Store the frozen cells in the -80℃ freezer.</br></br></p> | ||
− | + | </div> | |
− | + | <div id="list-item-4"> | |
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | |||
− | |||
− | |||
</br> | </br> | ||
<div class="row"> | <div class="row"> | ||
− | <a class="btn col-md-12" data-toggle="collapse" href="# | + | <a class="btn col-md-12" data-toggle="collapse" href="#Plasmid_Extraction" role="button" |
aria-expanded="false" aria-controls="multiCollapseExample1"> | aria-expanded="false" aria-controls="multiCollapseExample1"> | ||
− | + | Plasmid Extraction | |
<i class="fa fa-arrow-down fa-10" aria-hidden="true"></i> | <i class="fa fa-arrow-down fa-10" aria-hidden="true"></i> | ||
</a> | </a> | ||
</div> | </div> | ||
− | <div class="collapse multi-collapse" id=" | + | <div class="collapse multi-collapse" id="Plasmid_Extraction"> |
− | <p class="pcontent"></br></br> | + | <p class="pcontent"></br></br>1. Transfer 1.4 ml of well-grown bacterial culture to |
− | + | a centrifuge tube.</br></br> | |
− | + | 2. Centrifuge the tube at 16,000 xg for 1 minute to pellet the cells and | |
− | + | discard the supernatant completely.</br></br> | |
− | + | 3. Add 200 µl of FAPD1 Buffer (RNaseA added) to the cell pellet and resuspend | |
− | + | the cells completely by pipetting.</br></br> | |
− | + | ・ Make sure that RNaseA has been added into FAPD1 Buffer when first use.</br></br> | |
− | + | ・ No cell pellet should be visible after resuspension of the cells.</br></br> | |
− | 5. | + | 4. Add 200 µl of FAPD2 Buffer and gently invert the tube 5 ~ 10 times. Incubate |
− | + | the sample mixture at room temperature for 2 ~ 5 minutes to lyse the cells.</br></br> | |
− | 6. | + | ・ Do not vortex, vortex may shear genomic DNA. If necessary, continue inverting |
− | 7. | + | the tube until the lysate become clear.</br></br> ・ Make sure the tube transfer |
− | 8. | + | to clarify from turbid</br></br> |
− | + | ・ Do not proceed with the incubation over 5 minutes.</br></br> | |
− | + | 5. Add 300 µl of FAPD3 Buffer and invert the tube 5 ~ 10 times immediately to | |
− | + | neutralize the lysate.</br></br> | |
− | + | ・ Invert immediately after adding FAPD3 Buffer will avoid asymmetric | |
+ | precipitation.</br></br> | ||
+ | 6. Centrifuge at 16,000 xg for 3 minutess. to clarify the lysate. During | ||
+ | centrifugation, place a FAPD Column in a Collection Tube.</br></br> | ||
+ | 7. Transfer the supernatant carefully to the FAPD Column and centrifuge at | ||
+ | 16,000 xg for 1 minute. Discard the flow-through and place the column back to | ||
+ | the Collection Tube.</br></br> | ||
+ | ・ Do not transfer any white pellet into the column.</br></br> | ||
+ | 8. Add 400 µl of W1 Buffer to the FAPD Column and centrifuge at 16,000 xg for 1 | ||
+ | minute. Discard the flow-through and place the column back to the Collection | ||
+ | Tube.</br></br> | ||
+ | 9. Add 600 µl of Wash Buffer to the FAPD Column and centrifuge at 16,000 xg for | ||
+ | 1 minute. Discard the flow-through and place the column back to the Collection | ||
+ | Tube.</br></br> | ||
+ | ・ Make sure that ethanol (96 ~ 100 %) has been added into Wash Buffer when | ||
+ | first use.</br></br> | ||
+ | 10. Centrifuge at 16,000 xg for an additional 3 minutes to dry the FAPD Column.</br></br> | ||
+ | ・ Important step! The residual liquid should be removed thoroughly on this | ||
+ | step.</br></br> | ||
+ | 11. Place the FAPD Column to a new 1.5 ml microcentrifuge tube.</br></br> | ||
+ | 12. Add 30 µl of Elution Buffer or ddH2O to the membrane center of the FAPD | ||
+ | Column. Stand the column for 3 minute.</br></br> | ||
+ | ・ Important step! For effective elution, make sure that the elution solution is | ||
+ | dispensed on the membrane center and is absorbed completely.</br></br> | ||
+ | ・ Do not elute the DNA using less than suggested volume (50 µl). It will lower | ||
+ | the final yield.</br></br> | ||
+ | 13. Centrifuge at 16,000 xg for 3 minute to elute plasmid DNA and store the DNA | ||
+ | at -20 ℃.</br></br></p> | ||
− | |||
− | |||
− | + | </div> | |
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
+ | </div> | ||
+ | <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> | ||
− | <div | + | <div class="collapse multi-collapse" id="DNS"> |
− | + | ||
</br> | </br> | ||
− | < | + | <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> | |
− | + | <div class="col-6"> | |
− | + | <img class="contentimg" src="https://static.igem.org/mediawiki/2018/3/3e/T--NCKU_Tainan--home_xylose_new.jpg"> | |
− | + | </div> | |
− | + | <div class="col-6"> | |
− | + | <img class="contentimg" src="https://static.igem.org/mediawiki/2018/a/aa/T--NCKU_Tainan--home_glucose.png"></br></br> | |
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
</div> | </div> | ||
+ | <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> | ||
Line 481: | Line 482: | ||
</div> | </div> | ||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
<script> | <script> | ||
$(document).ready(function () { | $(document).ready(function () { | ||
$(window).scroll(function () { | $(window).scroll(function () { | ||
− | if ($(this).scrollTop() >= | + | if ($(this).scrollTop() >= 800) { |
var position = $("#sidelist").position(); | var position = $("#sidelist").position(); | ||
if (position == undefined) {} else { | if (position == undefined) {} else { | ||
Line 807: | Line 510: | ||
}); | }); | ||
</script> | </script> | ||
− | |||
− | |||
− | |||
− | |||
+ | <script src="https://2017.igem.org/Team:NCKU_Tainan/js/frame/T--NCKU_Tainan--jquery-1_12_4_min_js?action=raw&ctype=text/javascript"></script> | ||
+ | <script src="https://2018.igem.org/Template:NCKU_Tainan/js/bootstrap_min_js?action=raw&ctype=text/javascript"></script> | ||
+ | </div> | ||
</body> | </body> | ||
</html> | </html> | ||
− | |||
{{NCKU_Tainan/footer}} | {{NCKU_Tainan/footer}} |
Revision as of 10:42, 27 September 2018