Team:UNIPV-Pavia/Material Methods/Measurements/Tecan/test16settembre

From 2010.igem.org

(Difference between revisions)
(→EXPERIMENT DESCRIPTION)
 
(7 intermediate revisions not shown)
Line 14: Line 14:
==PLATE==
==PLATE==
<br>
<br>
-
<!--[[Image:UNIPV_Pavia_piastra27agosto.png| 620px | center ]]-->
+
[[Image:UNIPV_Pavia_piastra16settembre.png| 620px | center ]]
<br>
<br>
 +
==EXPERIMENT DESCRIPTION==
==EXPERIMENT DESCRIPTION==
===Motivation===
===Motivation===
Line 21: Line 22:
===Methods===
===Methods===
-
Inoculum (into 5 ml LB+Amp) from glycerol stock of:
+
Inoculum into 1 ml LB + a suitable antibiotic of 8 ul of glycerol stock of:
* <partinfo>pSB1A2</partinfo> - <partinfo>BBa_K300024</partinfo> (wiki name: I7)
* <partinfo>pSB1A2</partinfo> - <partinfo>BBa_K300024</partinfo> (wiki name: I7)
* <partinfo>pSB1A2</partinfo> - <partinfo>BBa_K300021</partinfo> (wiki name: I8)
* <partinfo>pSB1A2</partinfo> - <partinfo>BBa_K300021</partinfo> (wiki name: I8)
* <partinfo>pSB1A2</partinfo> - <partinfo>BBa_K300022</partinfo> (wiki name: I9)
* <partinfo>pSB1A2</partinfo> - <partinfo>BBa_K300022</partinfo> (wiki name: I9)
* <partinfo>pSB1A2</partinfo> - <partinfo>BBa_K300023</partinfo> (wiki name: I10)
* <partinfo>pSB1A2</partinfo> - <partinfo>BBa_K300023</partinfo> (wiki name: I10)
-
* <partinfo>pSB1A2</partinfo> - <partinfo>BBa_K300012</partinfo> (wiki name: I12)
+
* <partinfo>pSB1A2</partinfo> - <partinfo>BBa_K300019</partinfo> (wiki name: I12)
* <partinfo>pSB1A2</partinfo> - <partinfo>K173001</partinfo> (positive control)
* <partinfo>pSB1A2</partinfo> - <partinfo>K173001</partinfo> (positive control)
* <partinfo>pSB1A2</partinfo> - <partinfo>BBa_B0032</partinfo> (negative control)
* <partinfo>pSB1A2</partinfo> - <partinfo>BBa_B0032</partinfo> (negative control)
Line 36: Line 37:
* <partinfo>pSB4C5</partinfo> - <partinfo>BBa_K300027</partinfo> (wiki name I19) co-transformed with <partinfo>pSB1A3</partinfo> - <partinfo>BBa_T9002</partinfo>
* <partinfo>pSB4C5</partinfo> - <partinfo>BBa_K300027</partinfo> (wiki name I19) co-transformed with <partinfo>pSB1A3</partinfo> - <partinfo>BBa_T9002</partinfo>
* <partinfo>pSB1A3</partinfo> - <partinfo>BBa_T9002</partinfo> (negative control)
* <partinfo>pSB1A3</partinfo> - <partinfo>BBa_T9002</partinfo> (negative control)
 +
* <partinfo>pSB4C5</partinfo> - <partinfo>BBa_K300024</partinfo> (wiki name: I7-4C5)
 +
* <partinfo>pSB4C5</partinfo> - <partinfo>BBa_K300021</partinfo> (wiki name: I8-4C5)
 +
* <partinfo>pSB4C5</partinfo> - <partinfo>BBa_K300023</partinfo> (wiki name: I10-4C5)
 +
* <partinfo>pSB4C5</partinfo> - <partinfo>BBa_K300019</partinfo> (wiki name: I12-4C5)
They were let grow ON at +37°C, 220 rpm.
They were let grow ON at +37°C, 220 rpm.
-
The following day cultures were diluted 1:100 and let grow again for about five hours at +37°C, 220 rpm.
+
The following day cultures were diluted 1:100 in a final volume of 1 ml and let grow again for about five hours at +37°C, 220 rpm. Then cultures were centrifuged at 2000 rpm, 25°C, the supernatants were removed and the pellets were resuspended. Optical density (O.D.) of each cell culture was than measured with TECAN Infinte F200. Samples were diluted in order to obtain the same O.D. equal to 0,02. Then O.D. and green Fluorecence were measured.
 +
 
 +
==RESULTS==
 +
Cultures resulted immediately induced because of the high concentration of homoserine-lactone that remained in the solutions. This prevented from calculating the ODstart.
 +
 
 +
[[Image:UNIPV_Pavia_FluoVSODHC01609.png| 620px | center ]]
-
Optical density (O.D.) of each cell culture was than measured with TECAN Infinte F200. Samples were diluted in order to obtain the same O.D. equal to 0,02. Then O.D. and green Fluorecence were measured.
+
[[Image:UNIPV_Pavia_FluoVSODCo1609.png| 620px | center ]]
-
==RESULTS (?)==
+
[[Image:UNIPV_Pavia_FluoVSODLC01609.png| 620px | center ]]
<!-- table previous next test -->
<!-- table previous next test -->
Line 49: Line 59:
<table border="0" width="100%" class="menu">
<table border="0" width="100%" class="menu">
<tr>
<tr>
-
<td align="left" width="50%">[[Team:UNIPV-Pavia/Material Methods/Measurements/Tecan/test15settembre|Previous Test]]</td>
+
<td align="left" width="50%">[[Team:UNIPV-Pavia/Material Methods/Measurements/Tecan/test14settembre|Previous Test]]</td>
-
<td align="right" width="50%">[[Team:UNIPV-Pavia/Material Methods/Measurements/Tecan/test16settembre_bis|Next Test]]</td>
+
<td align="right" width="50%">[[Team:UNIPV-Pavia/Material Methods/Measurements/Tecan/test27settembre|Next Test]]</td>
</tr>
</tr>
</table>
</table>

Latest revision as of 21:27, 27 October 2010
Home
The Project
Materials & Methods
Parts
The Team
Notebook
Biological Safety
Pavia
Gallery
Sponsors

SEPTEMBER, 16TH


PLATE


UNIPV Pavia piastra16settembre.png


EXPERIMENT DESCRIPTION

Motivation

The experiment was performed to evaluate the initiation transcription point (in term of absorbance) for self-inducible devices.

Methods

Inoculum into 1 ml LB + a suitable antibiotic of 8 ul of glycerol stock of:

  • <partinfo>pSB1A2</partinfo> - <partinfo>BBa_K300024</partinfo> (wiki name: I7)
  • <partinfo>pSB1A2</partinfo> - <partinfo>BBa_K300021</partinfo> (wiki name: I8)
  • <partinfo>pSB1A2</partinfo> - <partinfo>BBa_K300022</partinfo> (wiki name: I9)
  • <partinfo>pSB1A2</partinfo> - <partinfo>BBa_K300023</partinfo> (wiki name: I10)
  • <partinfo>pSB1A2</partinfo> - <partinfo>BBa_K300019</partinfo> (wiki name: I12)
  • <partinfo>pSB1A2</partinfo> - <partinfo>K173001</partinfo> (positive control)
  • <partinfo>pSB1A2</partinfo> - <partinfo>BBa_B0032</partinfo> (negative control)
  • <partinfo>pSB4C5</partinfo> - <partinfo>BBa_K300030</partinfo> (wiki name I14) co-transformed with <partinfo>pSB1A3</partinfo> - <partinfo>BBa_T9002</partinfo>
  • <partinfo>pSB4C5</partinfo> - <partinfo>BBa_K300028</partinfo> (wiki name I15) co-transformed with <partinfo>pSB1A3</partinfo> - <partinfo>BBa_T9002</partinfo>
  • <partinfo>pSB4C5</partinfo> - <partinfo>BBa_K300029</partinfo> (wiki name I16) co-transformed with <partinfo>pSB1A3</partinfo> - <partinfo>BBa_T9002</partinfo>
  • <partinfo>pSB4C5</partinfo> - <partinfo>BBa_K300025</partinfo> (wiki name I17) co-transformed with <partinfo>pSB1A3</partinfo> - <partinfo>BBa_T9002</partinfo>
  • <partinfo>pSB4C5</partinfo> - <partinfo>BBa_K300026</partinfo> (wiki name I18) co-transformed with <partinfo>pSB1A3</partinfo> - <partinfo>BBa_T9002</partinfo>
  • <partinfo>pSB4C5</partinfo> - <partinfo>BBa_K300027</partinfo> (wiki name I19) co-transformed with <partinfo>pSB1A3</partinfo> - <partinfo>BBa_T9002</partinfo>
  • <partinfo>pSB1A3</partinfo> - <partinfo>BBa_T9002</partinfo> (negative control)
  • <partinfo>pSB4C5</partinfo> - <partinfo>BBa_K300024</partinfo> (wiki name: I7-4C5)
  • <partinfo>pSB4C5</partinfo> - <partinfo>BBa_K300021</partinfo> (wiki name: I8-4C5)
  • <partinfo>pSB4C5</partinfo> - <partinfo>BBa_K300023</partinfo> (wiki name: I10-4C5)
  • <partinfo>pSB4C5</partinfo> - <partinfo>BBa_K300019</partinfo> (wiki name: I12-4C5)

They were let grow ON at +37°C, 220 rpm.

The following day cultures were diluted 1:100 in a final volume of 1 ml and let grow again for about five hours at +37°C, 220 rpm. Then cultures were centrifuged at 2000 rpm, 25°C, the supernatants were removed and the pellets were resuspended. Optical density (O.D.) of each cell culture was than measured with TECAN Infinte F200. Samples were diluted in order to obtain the same O.D. equal to 0,02. Then O.D. and green Fluorecence were measured.

RESULTS

Cultures resulted immediately induced because of the high concentration of homoserine-lactone that remained in the solutions. This prevented from calculating the ODstart.

UNIPV Pavia FluoVSODHC01609.png
UNIPV Pavia FluoVSODCo1609.png
UNIPV Pavia FluoVSODLC01609.png



Previous Test Next Test

Tecan Test

Retrieved from "http://2010.igem.org/Team:UNIPV-Pavia/Material_Methods/Measurements/Tecan/test16settembre"