Difference between revisions of "Team:SSHS-Shenzhen/InterLab"
| Line 127: | Line 127: | ||
8）Mix A5 by pipetting up and down 3x and transfer 100 μl into A6... <br> | 8）Mix A5 by pipetting up and down 3x and transfer 100 μl into A6... <br> | ||
9）Mix A6 by pipetting up and down 3x and transfer 100 μl into A7... <br> | 9）Mix A6 by pipetting up and down 3x and transfer 100 μl into A7... <br> | ||
| − | 10）Mix A7 by pipetting up and down 3x and transfer 100 μl into A8... | + | 10）Mix A7 by pipetting up and down 3x and transfer 100 μl into A8... <br> |
11）Mix A8 by pipetting up and down 3x and transfer 100 μl into A9... <br> | 11）Mix A8 by pipetting up and down 3x and transfer 100 μl into A9... <br> | ||
12）Mix A9 by pipetting up and down 3x and transfer 100 μl into A10... <br> | 12）Mix A9 by pipetting up and down 3x and transfer 100 μl into A10... <br> | ||
13）Mix A10 by pipetting up and down 3x and transfer 100 μl into A11... <br> | 13）Mix A10 by pipetting up and down 3x and transfer 100 μl into A11... <br> | ||
| − | 14）Mix A11 by pipetting up and down 3x and transfer 100 μl into <b>liquid waste</b> <br> | + | 14）Mix A11 by pipetting up and down 3x and transfer 100 μl into <b>liquid waste</b> |
| + | |||
| + | <h2> | ||
| + | Monday, July 23, 2018 9:00(pm) | ||
| + | </h2> | ||
| + | <h3> | ||
| + | Specific data from our experiment | ||
| + | </h3> | ||
| + | <h2> | ||
| + | Note: we need to be aware of Re-mixing each of the row immediately before putting the plate into the plate reader. | ||
| + | </h2> | ||
| + | |||
| + | <h1> | ||
| + | 7/23:>Calibration 3: Fluorescence standard curve | ||
| + | </h1> | ||
| + | <h2> | ||
| + | Monday, July 23, 2018 2:00 | ||
| + | </h2> | ||
| + | <p id="para"> | ||
| + | 1) Spinning down fluorescein kit tube. <br> | ||
| + | 2) Prepare 10x fluorescein stock solution (100 μM) by resuspending fluorescein in 1 mL of 1xPBS. <br> | ||
| + | 3) Dilute the 10x fluorescein stock solution with 1xPBS to make a 1x fluorescein solution with concentration 10 μM: 100 μL of 10x fluorescein stock into 900 μL 1x PBS | ||
| + | </p> | ||
| + | |||
| + | <h2> | ||
| + | Monday, July 23, 2018 2:20 | ||
| + | </h2> | ||
| + | |||
| + | <p id="para"> | ||
| + | 1) Add 100 μl of PBS into wells A2, B2, C2, D2....A12, B12, C12, D12 <br> | ||
| + | 2) Add 200 μl of fluorescein 1x stock solution into A1, B1, C1, D1 <br> | ||
| + | 3) Transfer 100 μl of fluorescein stock solution from A1 into A2. <br> | ||
| + | 4) Mix A2 by pipetting up and down 3x and transfer 100 μl into A3… <br> | ||
| + | 5) Mix A3 by pipetting up and down 3x and transfer 100 μl into A4... <br> | ||
| + | 6) Mix A4 by pipetting up and down 3x and transfer 100 μl into A5... <br> | ||
| + | 7) Mix A5 by pipetting up and down 3x and transfer 100 μl into A6... <br> | ||
| + | 8) Mix A6 by pipetting up and down 3x and transfer 100 μl into A7... <br> | ||
| + | 9) Mix A7 by pipetting up and down 3x and transfer 100 μl into A8... <br> | ||
| + | 10) Mix A8 by pipetting up and down 3x and transfer 100 μl into A9... <br> | ||
| + | 11) Mix A9 by pipetting up and down 3x and transfer 100 μl into A10... <br> | ||
| + | 12) Mix A10 by pipetting up and down 3x and transfer 100 μl into A11... <br> | ||
| + | 13) Mix A11 by pipetting up and down 3x and transfer 100 μl into liquid waste | ||
| + | </p> | ||
| + | |||
| + | <h2> | ||
| + | Monday, July 23, 2018 3:00 | ||
| + | </h2> | ||
| + | <p id="para"> | ||
| + | 1) Repeat dilution series for rows B, C, D <br> | ||
| + | 2) Measure fluorescence of all samples in instrument <br> | ||
| + | 3) Record the data in our notebook <br> | ||
| + | 4) Import data into Excel sheet | ||
| + | </p> | ||
| + | |||
| + | <h1> | ||
| + | Cell measurement | ||
| + | </h1> | ||
| + | <h1> | ||
| + | 7/20:>Preparations | ||
| + | </h1> | ||
| + | <h2> | ||
| + | Friday July 18, 2018 9:00 | ||
| + | </h2> | ||
| + | |||
| + | <h3> | ||
| + | transform Escherichia coli DH5α with these following plasmids (all in pSB1C3): | ||
| + | </h3> | ||
| + | |||
<br><br><br><br><br> | <br><br><br><br><br> | ||
</body> | </body> | ||
Revision as of 13:16, 2 October 2018
Monday July 23, 2018 11:30
7/23: > Calibration 1: OD600 Reference point
Monday, July 23, 2018 9:00
1) Add 100 μl LUDOX into wells A1, B1, C1, D1
2) Add 100 μl of dd H2O into wells A2, B2, C2, D2
3) Measure absorbance at 600 nm of all samples in the measurement mode we plan to use for cell measurements
4) Record the data in both the table below and our notebook
5) Import data into Excel sheet provided
Monday, July 23, 2018 9:25(pm)
Specific data from our experiment
| LUDOX CL-X | H2O | |
| Replicate 1 | 0.058 | 0.038 |
| Replicate 2 | 0.059 | 0.037 |
| Replicate 3 | 0.053 | 0.037 |
| Replicate 4 | 0.055 | 0.036 |
| Arith. Mean | 0.056 | 0.037 |
| Corrected Abs600 | 0.019 | |
| Reference OD600 | 0.063 | |
| OD600/Abs600 | 3.238 |
Note: Carefully pipet liquids into the wells of 96 well plates in order to prevent the case of liquid sticking on the wall of wells and caused inaccurate data.
7/23:> Calibration 2: Particle Standard Curve
Monday, July 23, 2018 10:30
1) Obtain the tube labeled “Silica Beads” from the InterLab test kit and vortex 4 vigorously for 30 seconds.
2) Immediately pipet 96 μL microspheres into a 1.5 mL eppendorf tube
3) Add 904 μL of ddH2O to the microspheres
Monday, July 23, 2018 10:40
1）Add 100 μl of ddH2O into wells A2, B2, C2, D2....A12, B12, C12, D12
2）Vortex the tube containing the stock solution of microspheres vigorously for 10 seconds
3）Immediately add 200 μl of microspheres stock solution into A1
4）Transfer 100 μl of microsphere stock solution from A1 into A2.
5）Mix A2 by pipetting up and down 3x and transfer 100 μl into A3…
6）Mix A3 by pipetting up and down 3x and transfer 100 μl into A4...
7）Mix A4 by pipetting up and down 3x and transfer 100 μl into A5...
8）Mix A5 by pipetting up and down 3x and transfer 100 μl into A6...
9）Mix A6 by pipetting up and down 3x and transfer 100 μl into A7...
10）Mix A7 by pipetting up and down 3x and transfer 100 μl into A8...
11）Mix A8 by pipetting up and down 3x and transfer 100 μl into A9...
12）Mix A9 by pipetting up and down 3x and transfer 100 μl into A10...
13）Mix A10 by pipetting up and down 3x and transfer 100 μl into A11...
14）Mix A11 by pipetting up and down 3x and transfer 100 μl into liquid waste
Monday, July 23, 2018 9:00(pm)
Specific data from our experiment
Note: we need to be aware of Re-mixing each of the row immediately before putting the plate into the plate reader.
7/23:>Calibration 3: Fluorescence standard curve
Monday, July 23, 2018 2:00
1) Spinning down fluorescein kit tube.
2) Prepare 10x fluorescein stock solution (100 μM) by resuspending fluorescein in 1 mL of 1xPBS.
3) Dilute the 10x fluorescein stock solution with 1xPBS to make a 1x fluorescein solution with concentration 10 μM: 100 μL of 10x fluorescein stock into 900 μL 1x PBS
Monday, July 23, 2018 2:20
1) Add 100 μl of PBS into wells A2, B2, C2, D2....A12, B12, C12, D12
2) Add 200 μl of fluorescein 1x stock solution into A1, B1, C1, D1
3) Transfer 100 μl of fluorescein stock solution from A1 into A2.
4) Mix A2 by pipetting up and down 3x and transfer 100 μl into A3…
5) Mix A3 by pipetting up and down 3x and transfer 100 μl into A4...
6) Mix A4 by pipetting up and down 3x and transfer 100 μl into A5...
7) Mix A5 by pipetting up and down 3x and transfer 100 μl into A6...
8) Mix A6 by pipetting up and down 3x and transfer 100 μl into A7...
9) Mix A7 by pipetting up and down 3x and transfer 100 μl into A8...
10) Mix A8 by pipetting up and down 3x and transfer 100 μl into A9...
11) Mix A9 by pipetting up and down 3x and transfer 100 μl into A10...
12) Mix A10 by pipetting up and down 3x and transfer 100 μl into A11...
13) Mix A11 by pipetting up and down 3x and transfer 100 μl into liquid waste
Monday, July 23, 2018 3:00
1) Repeat dilution series for rows B, C, D
2) Measure fluorescence of all samples in instrument
3) Record the data in our notebook
4) Import data into Excel sheet