IGEM:Arizona State/2014/Notebook/asu igem 2014/2014/08/06

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(Autocreate 2014/08/06 Entry for IGEM:Arizona_State/2014/Notebook/asu_igem_2014)
Current revision (23:51, 6 August 2014) (view source)
(Entry title)
 
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==Entry title==
==Entry title==
-
* Insert content here...
+
* 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.
 +
 
 +
<u>Ethanol:</u>
 +
 
 +
*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
 +
 
 +
 
 +
{| {{table}} border="1" class="wikitable" style="text-align:center; width:400px; height:200px;"<!-- Ethanol Standard Table -->
 +
|+Ethanol Standard
 +
|- 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
 +
|-
 +
 
 +
|}
 +
 
 +
{| {{table}} border="1" class="wikitable" style="text-align:center; width:400px; height:200px;"<!--  Pyruvate Standard Table -->
 +
|+Pyruvate Standard
 +
|- 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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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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