Line 114: | Line 114: | ||
<div class="box fade-in two"> | <div class="box fade-in two"> | ||
<h1>MEETUP WITH TEAM KOREA_U_SEOUL</h1> | <h1>MEETUP WITH TEAM KOREA_U_SEOUL</h1> | ||
− | <h2>We had a brief, but productive meeting with team Korea_U_Seoul to discuss about what each team aimed to accomplish. We shared different ideas and comments about the progress each team has made. </h2><br><br> | + | <h2>We had a brief, but productive meeting with team Korea_U_Seoul to discuss about what each team aimed to accomplish. We shared different ideas and comments about the progress each team has made. </h2><br><br><br><br> |
<h1>Collaboration with iGEM BGU team</h1> | <h1>Collaboration with iGEM BGU team</h1> | ||
Line 121: | Line 121: | ||
<h2> | <h2> | ||
Terephthalic acid detection protocol Dissolve Terephtalic acid (TA) in water with strong base (Sodium hydroxide (NaOH)) in 1:2 concentration ratios. | Terephthalic acid detection protocol Dissolve Terephtalic acid (TA) in water with strong base (Sodium hydroxide (NaOH)) in 1:2 concentration ratios. | ||
− | Prepare a stock solution of 50 mL, 20 mM TA by adding: <br> | + | Prepare a stock solution of 50 mL, 20 mM TA by adding: <br><br> |
0.166 gr of TA <br> | 0.166 gr of TA <br> | ||
2.0 mL of 1M NaOH solution <br> | 2.0 mL of 1M NaOH solution <br> | ||
− | 48 mL double distilled water (DDW) < | + | 48 mL double distilled water (DDW) <br><br> |
Vortex for five minutes until the TA powder is completely dissolved. Titrate the solution with 1M HCl solution to achieve a pH of 7.5. <br> | Vortex for five minutes until the TA powder is completely dissolved. Titrate the solution with 1M HCl solution to achieve a pH of 7.5. <br> | ||
For a calibration curve: <br> | For a calibration curve: <br> | ||
− | Prepare 10 mL size batches of different TA concentrations. Dilute the initial TA stock solution with DDW to the following final concentrations: <br> | + | Prepare 10 mL size batches of different TA concentrations. Dilute the initial TA stock solution with DDW to the following final concentrations: <br><br> |
− | -15mM, 10mM, 5mM, 1mM, 0.5mM, 0.1mM, 0.05mM, 0.01mM, 0.005mM, 0.001mM. <br> | + | -15mM, 10mM, 5mM, 1mM, 0.5mM, 0.1mM, 0.05mM, 0.01mM, 0.005mM, 0.001mM. <br><br> |
Pipette 400 μL of each concentration in a non-UV absorbent 96 wells microtiter plate. Make triplicates of each concentration. <br> | Pipette 400 μL of each concentration in a non-UV absorbent 96 wells microtiter plate. Make triplicates of each concentration. <br> | ||
− | For baseline readings, measure florescence in a plate reader in the following values: <br> | + | For baseline readings, measure florescence in a plate reader in the following values: <br><br> |
-Excitation: 314 nm. <br> | -Excitation: 314 nm. <br> | ||
-Emission: 426 nm. <br> | -Emission: 426 nm. <br> | ||
-Gain: 100 HZ to detect high concentration and 200 Hz to detect small concentration <br> | -Gain: 100 HZ to detect high concentration and 200 Hz to detect small concentration <br> | ||
− | -Number of Flashes: 150 to detect high concentration and 200 to detect small concentration<br> | + | -Number of Flashes: 150 to detect high concentration and 200 to detect small concentration<br><br> |
Following the measurements irradiate the samples with UV light (365 nm) for one hour using a UV transilluminaitor.<br> | Following the measurements irradiate the samples with UV light (365 nm) for one hour using a UV transilluminaitor.<br> | ||
− | <br>Repeat the florescence measurements using the plate reader. (If needed, prolong the exposure time of the sample to enhance florescence). Create a calibration curve from the fluorescence values that were received for each concentration. Use the curve to extract the concentrations of TA in a sample in unknown concentrations. | + | <br>Repeat the florescence measurements using the plate reader. (If needed, prolong the exposure time of the sample to enhance florescence). Create a calibration curve from the fluorescence values that were received for each concentration. Use the curve to extract the concentrations of TA in a sample in unknown concentrations.<br><br> |
</h2> | </h2> | ||
</div> | </div> |
Revision as of 15:54, 16 October 2016