IGEM:Arizona State/2014/Notebook/asu igem 2014/2014/08/06: Difference between revisions

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:::-Flask 3: 3 g/L pyruvate (.09 g)(no glucose)
:::-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)
**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
<u>Ethanol Standard</u>
{| {{table}} border="1" align=center <!-- Ethanol Standard Table -->
|- valign="top"
| bgcolor=#cfcfcf | Desired Concentration (mol/L)
| bgcolor=#cfcfcf | Water Added (ml)
| bgcolor=#cfcfcf | Pure Ethanol Added (ml)
|-
| bgcolor=#cfcfcf | 1.13
| bgcolor=#cfcfcf | 28.021
| bgcolor=#cfcfcf | 1.979
|-
|| .757|| .33  || .620
|-
|| .38 || .664  || .336
|-
|| .3|| .735 || .265
|-
|| .2 || .823 || .177
|-
|| .1 || .912 || .088
|-
|}
<u>Pyruvate Standard</u>
{| {{table}} border="1" align=center <!--  Pyruvate Standard Table -->
|- valign="top"
| bgcolor=#cfcfcf | Desired Concentration (g/L)
| bgcolor=#cfcfcf | Water Added (ml)
| bgcolor=#cfcfcf | Pyruvate Added (ml)
|-
| bgcolor=#cfcfcf | 3.0
| bgcolor=#cfcfcf | 30 ml
| bgcolor=#cfcfcf | .09 g
|-
|| 2.0 || .333  || .667
|-
|| 1.0 || .666  || .333
|-
|| .7 || .767 || .233
|-
|| .5 || .833 || .167
|-
|| .3 || .900 || .100
|-
|}


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Revision as of 20:31, 6 August 2014

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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)
1.13 28.021 1.979
.757 .33 .620
.38 .664 .336
.3 .735 .265
.2 .823 .177
.1 .912 .088

Pyruvate Standard

Desired Concentration (g/L) Water Added (ml) Pyruvate Added (ml)
3.0 30 ml .09 g
2.0 .333 .667
1.0 .666 .333
.7 .767 .233
.5 .833 .167
.3 .900 .100


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