Sunday, October 17, 2010

the 3rd day @ 2:10 PM

Innoculation time…

After waiting for 2 days for the fermenter to be fully set up, today Mr Shaman shows us how to do inoculation and take sample aseptically. Now is time to inoculate our lovely wild type yeast, Saccharomyces cerevisiae into the fermenter and let them grow.

But before inoculate the culture we need to set the parameter required by the yeast at the fermenter.

-    Aeration rate: 1L /minute
-    Speed: 700 rpm
-    Calibrate pH
     ·    1st calibration for 4.01
     ·    2nd calibration for 7.01
-    Temperature
-    Calibrate the oxygen
     ·    Sparge the Nitrogen gas into the vessel – to remove the out O2 in the media. Then stop adding N2 when the PO2 data is stable and indicate as 0%.
     ·    Start aeration to increase the O2 level in the media and let the data of PO2 stable. Then indicate it is as 100 %.
-    Antifoam
-    Base line


Here are several important steps that we need to follow:
1.    Cut off the aeration while inoculate the sample
2.    Spray the inoculation inlet with 70% ethanol
3.    Open inlet probe and then spray again
4.    Spray the end of tube at conical flask containing our culture with 70% ethanol.
5.    Quickly place the tube to the inlet probe.
6.    Start aeration again after finish inoculation. The reason is to increase the pressure in the vessel so that we can reduce the risk of contamination.


Sampling time...




Let me explain how to do sampling without taking out the sample bottle. It is because we want to reduce the risk of contamination.
1.    Put syringe at the filter.
2.    Unclamp the sample line A
3.    Put back on syringe then the sample will flow out into the sampling bottle.
4.    Reclamp the sample line.
5.    Unclamp the sample line B
6.    Spray the end tube of sample line B with 70% ethanol.
7.    Push the syringe and the sample will flow into the test tube.
8.    The sample now ready to be observed
9.    Don’t forget to reclamp the sample line B
10.    Spray again the end tube and wrap it with cotton and aluminum foil.
We need to do sampling from 12.30 for first reading at time 0. After that we need to sample out to take OD600 value and glucose reading every hour from 2pm until 11 am on the next day.

Sunday, October 17, 2010

the 3rd day @ 2:10 PM

Innoculation time…

After waiting for 2 days for the fermenter to be fully set up, today Mr Shaman shows us how to do inoculation and take sample aseptically. Now is time to inoculate our lovely wild type yeast, Saccharomyces cerevisiae into the fermenter and let them grow.

But before inoculate the culture we need to set the parameter required by the yeast at the fermenter.

-    Aeration rate: 1L /minute
-    Speed: 700 rpm
-    Calibrate pH
     ·    1st calibration for 4.01
     ·    2nd calibration for 7.01
-    Temperature
-    Calibrate the oxygen
     ·    Sparge the Nitrogen gas into the vessel – to remove the out O2 in the media. Then stop adding N2 when the PO2 data is stable and indicate as 0%.
     ·    Start aeration to increase the O2 level in the media and let the data of PO2 stable. Then indicate it is as 100 %.
-    Antifoam
-    Base line


Here are several important steps that we need to follow:
1.    Cut off the aeration while inoculate the sample
2.    Spray the inoculation inlet with 70% ethanol
3.    Open inlet probe and then spray again
4.    Spray the end of tube at conical flask containing our culture with 70% ethanol.
5.    Quickly place the tube to the inlet probe.
6.    Start aeration again after finish inoculation. The reason is to increase the pressure in the vessel so that we can reduce the risk of contamination.


Sampling time...




Let me explain how to do sampling without taking out the sample bottle. It is because we want to reduce the risk of contamination.
1.    Put syringe at the filter.
2.    Unclamp the sample line A
3.    Put back on syringe then the sample will flow out into the sampling bottle.
4.    Reclamp the sample line.
5.    Unclamp the sample line B
6.    Spray the end tube of sample line B with 70% ethanol.
7.    Push the syringe and the sample will flow into the test tube.
8.    The sample now ready to be observed
9.    Don’t forget to reclamp the sample line B
10.    Spray again the end tube and wrap it with cotton and aluminum foil.
We need to do sampling from 12.30 for first reading at time 0. After that we need to sample out to take OD600 value and glucose reading every hour from 2pm until 11 am on the next day.