Difference between revisions of "Team:Kyoto/SpecialMethods"
| Line 98: | Line 98: | ||
| − | <h5 id="Boil Method"><font face="Segoe UI">1)Boil Method</font></ | + | <h5 id="Boil Method"><font face="Segoe UI">1)Boil Method</font></h5> |
<p><font size="2px">1.Monitor OD of yeast and incubate until it becomes OD≒1. | <p><font size="2px">1.Monitor OD of yeast and incubate until it becomes OD≒1. | ||
<br>2.Centrifuge yeast at 3500 rpm for 5 min | <br>2.Centrifuge yeast at 3500 rpm for 5 min | ||
Revision as of 22:08, 17 October 2018
1)Boil Method
1.Monitor OD of yeast and incubate until it becomes OD≒1.
2.Centrifuge yeast at 3500 rpm for 5 min
3.Discard the supernatant
4.Pipette the culture residual substance and transfer 1 ml of it to an Eppendorf tube
5.Centrifuge on FLASH and then decantate it
6.Add 1 ml of distilled water
7.Vortex
8.Centrifuge yeast at FLASH
9.Discard the supernatant (wash 1st time)
10.Repeat steps 6-9 (wash 2nd)
11.Repeat steps 6-9 (wash 3rd)
12.Cryopreservation
13.Add 1 ml of distilled water and vortex
14.Boil it with hot water for 10 min
· Fit the tube in the sponge and put it in boiling water
(· Keep the fire between low to medium heat)
15.Centrifuge at 10000 rpm for 5 min, and remove 900μl of the supernatant
16.Measure Na+ concentration(Atomic Absorption Spectrometry)
picture here
2)
picture here
3)
picture here
4)
picture here