Team:Kyoto/SpecialMethods

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Table of contents

• 1)Boil Method
• 2)
• 3)
• 4)

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)

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4)

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