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Entry title
- Checked plates from 8/5
- Only + control on chloro and RBS-TesA on cholro had growth.
- Obtained acc plasmid (chloro resistance). Started plate and liquid colonies.
Ethanol:
- At 8:30 AM, O.D. of cells was approx. 1.2
- Resuspended in 30 ml PBS, adjusted O.D. to match at 1.0, resuspended in PBS again
- Began resting cell assay (9:15 AM):
- -Flask 1: 1 g/L pyruvate (.03 g)
- -Flask 2: 3 g/L pyruvate (.09 g)+ 1 g/L glucose (.03 g)
- -Flask 3: 3 g/L pyruvate (.09 g)(no glucose)
- Flasks were kept in 32°C shaker/incubator with 1 ml samples taken for Hrs. 1,2,3,6, and 8. (24 Hr. sample will be taken tomorrow)
- Made standards of ethanol and pyruvate to be run on the HPLC:
- Standards were first made of the max possible amount of each substance present in the system. (in 30 ml) These concentrations were then used as a stock to be diluted for subsequent concentrations
Ethanol Standard
| Desired Concentration (mol/L)
|
Water Added (ml)
|
Pure Ethanol Added (ml)
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| 1.13
|
28.021
|
1.979
|
| .757 |
.33 |
.620
|
| .38 |
.664 |
.336
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| .3 |
.735 |
.265
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| .2 |
.823 |
.177
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| .1 |
.912 |
.088
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Pyruvate Standard
| Desired Concentration (g/L)
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Water Added (ml)
|
Pyruvate Added (ml)
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| 3.0
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30 ml
|
.09 g
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| 2.0 |
.333 |
.667
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| 1.0 |
.666 |
.333
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| .7 |
.767 |
.233
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| .5 |
.833 |
.167
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| .3 |
.900 |
.100
|
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