Team:TU Delft/27 July 2010 content

From 2010.igem.org

(Difference between revisions)
(→RBS Characterization)
(→Alkane degradation)
Line 2: Line 2:
[https://2010.igem.org/Team:TU_Delft#/blog?blog=26_July_2010 Yesterday's] digestion products were checked on a gel.
[https://2010.igem.org/Team:TU_Delft#/blog?blog=26_July_2010 Yesterday's] digestion products were checked on a gel.
[[Image:TUDelft_Kira_digestion_2010_07_27.png|500px|thumb|left|Digestion]]
[[Image:TUDelft_Kira_digestion_2010_07_27.png|500px|thumb|left|Digestion]]
 +
 +
{| style="color:black; background-color:white;" cellpadding="5" cellspacing="0" border="1"
 +
|'''#'''
 +
|'''Lane description'''
 +
|'''As expected?'''
 +
|-
 +
|1
 +
|[https://2010.igem.org/Image:TU_Delft_SmartLadder.jpg SmartLadder] (5μl)
 +
|
 +
|-
 +
|2
 +
|rubA3 (cut)
 +
|✓
 +
|-
 +
|3
 +
|rubA3 (uncut)
 +
|✓
 +
|-
 +
|4
 +
|rubA4 (cut)
 +
|✓
 +
|-
 +
|5
 +
|rubA4 (uncut)
 +
|✓
 +
|-
 +
|6
 +
|rubR (cut)
 +
|✓
 +
|-
 +
|7
 +
|rubR (uncut)
 +
|✓
 +
|-
 +
|8
 +
|ladA (cut)
 +
|✓
 +
|-
 +
|9
 +
|ladA (uncut)
 +
|✓
 +
|-
 +
|10
 +
|ADH (cut)
 +
|✓
 +
|-
 +
|11
 +
|ADH (uncut)
 +
|✓
 +
|-
 +
|12
 +
|ALDH (cut)
 +
|✓
 +
|-
 +
|13
 +
|ALDH (uncut)
 +
|✓
 +
|-
 +
|14
 +
|J61100 (cut)
 +
|✓
 +
|-
 +
|15
 +
|J61100 (uncut)
 +
|✓
 +
|-
 +
|16
 +
|J61101 (cut)
 +
|✓
 +
|-
 +
|17
 +
|J61101 (uncut)
 +
|✓
 +
|-
 +
|18
 +
|J61107 (cut)
 +
|✓
 +
|-
 +
|19
 +
|J61107 (uncut)
 +
|✓
 +
|}
 +
 +
 +
<html><br/></html>
<html><br/></html>
 +
==RBS Characterization==
==RBS Characterization==
For our RBS characterization project, 5 different RBS sequences from the [http://partsregistry.org/Ribosome_Binding_Sites/Prokaryotic/Constitutive/Anderson Anderson RBS family] where placed in front of GFP and measured over 18 hours using a Gen5 fluorenscence and absorbance plate reader
For our RBS characterization project, 5 different RBS sequences from the [http://partsregistry.org/Ribosome_Binding_Sites/Prokaryotic/Constitutive/Anderson Anderson RBS family] where placed in front of GFP and measured over 18 hours using a Gen5 fluorenscence and absorbance plate reader

Revision as of 07:32, 28 July 2010

Alkane degradation

Yesterday's digestion products were checked on a gel.

Digestion
# Lane description As expected?
1 SmartLadder (5μl)
2 rubA3 (cut) ✓
3 rubA3 (uncut) ✓
4 rubA4 (cut) ✓
5 rubA4 (uncut) ✓
6 rubR (cut) ✓
7 rubR (uncut) ✓
8 ladA (cut) ✓
9 ladA (uncut) ✓
10 ADH (cut) ✓
11 ADH (uncut) ✓
12 ALDH (cut) ✓
13 ALDH (uncut) ✓
14 J61100 (cut) ✓
15 J61100 (uncut) ✓
16 J61101 (cut) ✓
17 J61101 (uncut) ✓
18 J61107 (cut) ✓
19 J61107 (uncut) ✓




RBS Characterization

For our RBS characterization project, 5 different RBS sequences from the [http://partsregistry.org/Ribosome_Binding_Sites/Prokaryotic/Constitutive/Anderson Anderson RBS family] where placed in front of GFP and measured over 18 hours using a Gen5 fluorenscence and absorbance plate reader 26 07 2010 Rbs.png

Strength was calculated by taking the mean of the ratio between the expression of known RBS ([http://partsregistry.org/Part:BBa_B0032 B0032]) and expression Anderson RBS over some time. Expression being the measured fluorescence divided by measured biomass (absorbance, OD).

The measurements where taken from the part of the curve where optimal growth can be assumed, so from 0:40 until 2:30

RBS Strength
[http://partsregistry.org/Part:BBa_J61100 J61100] 0.047513
[http://partsregistry.org/Part:BBa_J61101 J61101] 0.119831
[http://partsregistry.org/Part:BBa_J61107 J61107] 0.065454
[http://partsregistry.org/Part:BBa_J61117 J61117] 0.038518
[http://partsregistry.org/Part:BBa_J61127 J61127] 0.087334
[http://partsregistry.org/Part:BBa_B0032 B0032] 0.300000

Next step: We want to relate the RBS sequence to the strength, so it might be possible to say predict something about the other Anderson RBS sequences.

Retrieved from "http://2010.igem.org/Team:TU_Delft/27_July_2010_content"