week8 summary

//It’s helpful if you first have salt, water, saliva, and a paper cup for trash.

Label 1.5m Eppendorf Tube with your name
Prepare Saline (=Salt) solution
Use ca. 5ml of water, mix with a pinch of salt
// A pinch of salt in about 5 mm of water in the bottom of a paper cup
Rinse your mouth
// for one minute
Transfer to 1.5ml Eppendorf Tube.
If you use a Paper Cup, bend it and fill the tube
// No need to use Pipetman.
Centrifuge at 4KG for 90sec
// bentolab’s UI is tricky and you should read the manual beforehand
Recover the Pellet by decanting the Supernatant
The Pellet is the white concentration of cells on the bottom of the tube. The liquid above is the Supernatant
Resuspend the Pellet
Mix the Pellet with the remaining Supernatant by repeatedly flipping and tapping it.
Transfer the sample to a 0.2ml PCR tube.
You can use a 2-20 or 10-100 Pipette.
Label the tube.
// It is useful to write your name on the lid of tube as well
Heat the Sample at 99ºC for 10min
Mix the Sample by flipping and tapping
Centrifuge at 8KG for 90sec
Use the Adapter Eppendorf tube to fit the PCR tubes in the Centrifuge
The DNA is now in the Supernatant, NOT in the Pellet
Transfer the Supernatant is a new PCR Tube
Use a 20uL Pipette to transfer the Supernatant - and only the Supernatant - to a new PCR Tube
Label and Freeze at -20ºC

17:00- slime mold second experiment with 桜木さん、土屋さん
Using a pill crusher, I grinded the oatmeal and drew patterns.

18:45- E.coli spreading out with inoculation loop

notes:

The reason for rubbing the medium with a loop, like frottage, is to create a gradient of bacterial concentration in the trail, making it easier to isolate colonies that will be formed by a single E.coli.
Loosen the lid of the tube containing E.coli to make it easier to open with one hand.
The bag of inoculation loop should be opened in the clean bench and left in there until discarded.
This time, we incubated at 30°C.

Written on April 28, 2022