Team:Imperial College London/Wetlab/Protocols/Autoinduction

From 2009.igem.org

(Difference between revisions)
 
(4 intermediate revisions not shown)
Line 1: Line 1:
{{Imperial/09/TemplateTop}}
{{Imperial/09/TemplateTop}}
{{Imperial/09/Tabs/Main/Wetlab/Protocols}}
{{Imperial/09/Tabs/Main/Wetlab/Protocols}}
-
 
{| style="color:#CCC; background-color:#325d97;" cellpadding="6" cellspacing="0" border="3"
{| style="color:#CCC; background-color:#325d97;" cellpadding="6" cellspacing="0" border="3"
! Section
! Section
! Assay
! Assay
! Overview and Aims
! Overview and Aims
-
 
|- style="color:#333; background-color:#CCCCFF;" cellpadding="6" cellspacing="0" border="1"
|- style="color:#333; background-color:#CCCCFF;" cellpadding="6" cellspacing="0" border="1"
 +
| <b>Promoter Characterisation</b>
| [[Team:Imperial_College_London/Wetlab/Protocols/SecondaryCarbon|Secondary Carbon Source Diauxie Growth]]
| [[Team:Imperial_College_London/Wetlab/Protocols/SecondaryCarbon|Secondary Carbon Source Diauxie Growth]]
|  
|  
Line 13: Line 12:
* To determine the best secondary carbon source for optimal growth
* To determine the best secondary carbon source for optimal growth
|- style="color:#333; background-color:#CCCCFF;" cellpadding="6" cellspacing="0" border="1"
|- style="color:#333; background-color:#CCCCFF;" cellpadding="6" cellspacing="0" border="1"
 +
| <b>Auto-Induction</b>
 +
| [[Team:Imperial_College_London/Wetlab/Protocols/Glucose delay|Glucose Time Delay]]
 +
| * Characterise the tunable time duration it takes before GFP expression (M2 activation)
|}
|}
-
 
-
 
-
 
-
 
-
 
-
 
-
 
-
 
-
 
-
 
-
 
-
 
-
 
-
 
-
 
-
 
-
 
-
 
-
==Glucose time delay==
 
-
===Aims===
 
-
 
-
* Characterise the tunable time duration it takes before GFP expression (M2 activation)
 
-
 
-
===Assay===
 
-
 
-
*The cells will be grown until OD= 0.7. 
 
-
 
-
*The RFP value will be monitered, although it is the GFP values that are more critical in this assay.
 
-
 
-
*OD and fluorescence data for GFP which can be converted in [http://partsregistry.org/cgi/measurement/new_batch.cgi Specific Promoter Units (SPUs)].
 
-
 
-
*The secondary carbon source will be taken from the previous experiment (see Secondary carbon source selection for CRP promoter)
 
-
 
-
*The IPTG concentrations will be taken from the previous experiment (see Determining concentration of IPTG)
 
-
 
-
[[Team:Imperial_College_London/Wetlab/Protocols/Glucose delay|See Glucose time delay for details]]
 
{{Imperial/09/TemplateBottom}}
{{Imperial/09/TemplateBottom}}

Latest revision as of 15:01, 23 September 2009



Section Assay Overview and Aims
Promoter Characterisation Secondary Carbon Source Diauxie Growth
  • To generate the diauxie growth curves, as well as normal growth curves for Top-10 for modelling
  • To determine the best secondary carbon source for optimal growth
Auto-Induction Glucose Time Delay * Characterise the tunable time duration it takes before GFP expression (M2 activation)

Mr. Gene   Geneart   Clontech   Giant Microbes