Difference between revisions of "Team:Kyoto/SpecialMethods"
SAKANAhuman (Talk | contribs) |
SAKANAhuman (Talk | contribs) |
||
| Line 136: | Line 136: | ||
<h5 id="Boil Method"><font face="Segoe UI">1)Boil Method</font></h2> | <h5 id="Boil Method"><font face="Segoe UI">1)Boil Method</font></h2> | ||
| − | <p> | + | <p> |
| + | 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) | ||
| + | </p> | ||
<img src="https://2018.igem.org"><p>picture here</p> | <img src="https://2018.igem.org"><p>picture here</p> | ||
<br><br><br> | <br><br><br> | ||
Revision as of 17:58, 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