Team:Stockholm/15 October 2010

From 2010.igem.org

(Difference between revisions)
(New page: {{Stockholm/Top2}} ==Andreas==)
(→Andreas)
Line 1: Line 1:
{{Stockholm/Top2}}
{{Stockholm/Top2}}
==Andreas==
==Andreas==
 +
===Growth curve assay===
 +
====Setup====
 +
Inoculated new flasks from ON cultures set 14/10:
 +
*250 ml E-flasks
 +
*25 ml LB + Amp 100 + 1 mM IPTG
 +
*250 μl ON culture
 +
*#BL21 pEX.SOD⋅His
 +
*#BL21 pEX.nTra10⋅SOD⋅His
 +
*#BL21 pEX.nTAT⋅SOD⋅His
 +
*#BL21 pEX.nLMWP⋅SOD⋅His
 +
*37 °C, 225 rpm
 +
 +
OD<sub>590</sub> measurements made every 30 min with appropriate dilution in fresh LB.
 +
 +
====OD<sub>590</sub> measurements====
 +
 +
{|border="1" cellpadding="1" cellspacing="0"
 +
|&nbsp;
 +
!width="50"|0 min
 +
!width="50"|30 min
 +
!width="50"|60 min
 +
!width="50"|90 min
 +
!width="50"|120 min
 +
!width="50"|150 min
 +
!width="50"|180 min
 +
!width="50"|210 min
 +
!width="50"|240 min
 +
!width="50"|270 min
 +
!width="50"|300 min
 +
!width="50"|330 min
 +
!width="50"|360 min
 +
!width="50"|390 min
 +
|-
 +
|'''pEX.SOD&sdot;His'''
 +
|align="center"|0.035
 +
|align="center"|0.047
 +
|align="center"|0.090
 +
|align="center"|0.180
 +
|align="center"|0.195
 +
|align="center"|0.176
 +
|align="center"|0.251
 +
|align="center"|0.177
 +
|align="center"|0.239
 +
|align="center"|0.140
 +
|align="center"|0.164
 +
|align="center"|0.205
 +
|align="center"|0.196
 +
|align="center"|0.193
 +
|-
 +
|'''pEX.nTra10&sdot;SOD&sdot;His'''
 +
|align="center"|0.037
 +
|align="center"|0.050
 +
|align="center"|0.092
 +
|align="center"|0.163
 +
|align="center"|0.156
 +
|align="center"|0.132
 +
|align="center"|0.175
 +
|align="center"|0.130
 +
|align="center"|0.164
 +
|align="center"|0.116
 +
|align="center"|0.155
 +
|align="center"|0.167
 +
|align="center"|0.154
 +
|align="center"|0.168
 +
|-
 +
|'''pEX.nTAT&sdot;SOD&sdot;His'''
 +
|align="center"|0.033
 +
|align="center"|0.046
 +
|align="center"|0.090
 +
|align="center"|0.170
 +
|align="center"|0.177
 +
|align="center"|0.161
 +
|align="center"|0.225
 +
|align="center"|0.153
 +
|align="center"|0.225
 +
|align="center"|<font style="color:red">0.045</font>
 +
|align="center"|0.170
 +
|align="center"|0.162
 +
|align="center"|0.193
 +
|align="center"|0.200
 +
|-
 +
|'''pEX.nLMWP&sdot;SOD&sdot;His'''
 +
|align="center"|0.031
 +
|align="center"|0.042
 +
|align="center"|0.86
 +
|align="center"|0.174
 +
|align="center"|0.192
 +
|align="center"|0.178
 +
|align="center"|0.237
 +
|align="center"|0.170
 +
|align="center"|0.242
 +
|align="center"|0.137
 +
|align="center"|0.180
 +
|align="center"|0.185
 +
|align="center"|0.200
 +
|align="center"|0.200
 +
|-
 +
|'''Dilution factor'''
 +
|align="center"|1
 +
|align="center"|1
 +
|align="center"|1
 +
|align="center"|1
 +
|align="center"|2
 +
|align="center"|4
 +
|align="center"|5
 +
|align="center"|10
 +
|align="center"|10
 +
|align="center"|20
 +
|align="center"|20
 +
|align="center"|20
 +
|align="center"|20
 +
|align="center"|20
 +
|}
 +
 +
[[image:Growth_assay_plot_15oct.png|200px|thumb|right|'''Line chart of plotted OD(590) measurements from growth assay.''']]
 +
 +
====Results====
 +
Results indicate no significant difference in growth between different cultures. Slightly lower curve for the nTra10-expressing clone, which is somewhat interesting, since a similar progress was seen in the canceled experiment from 14/10. At least one more experiment will be run next week, using new BL21 clones; this will show whether this is just a coincidence or not.

Revision as of 15:53, 15 October 2010

Home Project Idea Lab Work Results Modelling Team       Follow us on and        2010.igem.org

↳   Notebook Safety Protocols


Contents

Andreas

Growth curve assay

Setup

Inoculated new flasks from ON cultures set 14/10:

  • 250 ml E-flasks
  • 25 ml LB + Amp 100 + 1 mM IPTG
  • 250 μl ON culture
    1. BL21 pEX.SOD⋅His
    2. BL21 pEX.nTra10⋅SOD⋅His
    3. BL21 pEX.nTAT⋅SOD⋅His
    4. BL21 pEX.nLMWP⋅SOD⋅His
  • 37 °C, 225 rpm

OD590 measurements made every 30 min with appropriate dilution in fresh LB.

OD590 measurements

  0 min 30 min 60 min 90 min 120 min 150 min 180 min 210 min 240 min 270 min 300 min 330 min 360 min 390 min
pEX.SOD⋅His 0.035 0.047 0.090 0.180 0.195 0.176 0.251 0.177 0.239 0.140 0.164 0.205 0.196 0.193
pEX.nTra10⋅SOD⋅His 0.037 0.050 0.092 0.163 0.156 0.132 0.175 0.130 0.164 0.116 0.155 0.167 0.154 0.168
pEX.nTAT⋅SOD⋅His 0.033 0.046 0.090 0.170 0.177 0.161 0.225 0.153 0.225 0.045 0.170 0.162 0.193 0.200
pEX.nLMWP⋅SOD⋅His 0.031 0.042 0.86 0.174 0.192 0.178 0.237 0.170 0.242 0.137 0.180 0.185 0.200 0.200
Dilution factor 1 1 1 1 2 4 5 10 10 20 20 20 20 20
Line chart of plotted OD(590) measurements from growth assay.

Results

Results indicate no significant difference in growth between different cultures. Slightly lower curve for the nTra10-expressing clone, which is somewhat interesting, since a similar progress was seen in the canceled experiment from 14/10. At least one more experiment will be run next week, using new BL21 clones; this will show whether this is just a coincidence or not.

Retrieved from "http://2010.igem.org/Team:Stockholm/15_October_2010"