Revision as of 13:19, 2 October 2018

Monday July 23, 2018 11:30

7/23: > Calibration 1:​ OD​600​ 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

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 ddH​2​O​ 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

Cell measurement

7/20:>Preparations

Friday July 18, 2018 9:00

transform ​Escherichia coli DH5α with these following plasmids (all in pSB1C3):