IGEM:Cambridge/2008/Notebook/Turing Pattern Formation/2008/08/09: Difference between revisions

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==Result from Erythromycin test==
==Result from Erythromycin test==


Problem : colonies on each plates
Problem : colonies on each plate


*Reason : Erythromyci has to be diluted in ethanol!!!!
* Reason : Erythromycin has to be diluted in ethanol, not water
* To be sure, took a bacillus colony with no resistance and plated it onto Ery 0.5


We will do that again on monday.
On Monday we will remake the Erythromycin plates, and replate everything with the proper controls. Unfortunately, we do not have a positive control for erythromycin resistance, but if things die (i.e. negative control works), we should be in business.
 
==Plans for Monday==
===Lab Work===
*Be sure that the Spc50 plates are working, as the control for Spc50 grew and shouldn't have. My guess (Dan) is that we did not  create the stock correctly or used the wrong solvent to dilute it.
*Finish all the agr PCRs, but especially the RBS.
*Confirm that amyE insertion works, using the amylase/ery tests.
*Confirm that with ECE153 inserted,  we get detectable fluoresence in a microscope.
*Confirm that ECE166 transformation works, using plasmid miniprepping.
**Find the qiagen miniprep kit or adapt the protocol.
*Confirm that when ECE166 is transformed, we get detectable fluoresence in a microscope.
===Book/Computer Work===
*Find good promoters in vectors and order PCR primers to get them out
*Find out how to assay for/make AIP
**Find/Contact Jim Ajioka
 
==Longer Term Plans==
*Ligate the RBS into a BB vector so we can build a RBS/GFP construct.
*Put the RBS/GFP construct into ECE166 and ECE 153.  
*Compare the 'out of the box' ECE153 with ECE153 with RBS/GFP biobrick inserts
*Compare the 'out of the box' ECE166 with ECE166 with RBS/GFP biobrick inserts




Revision as of 16:35, 10 August 2008

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Result from Erythromycin test

Problem : colonies on each plate

  • Reason : Erythromycin has to be diluted in ethanol, not water
  • To be sure, took a bacillus colony with no resistance and plated it onto Ery 0.5

On Monday we will remake the Erythromycin plates, and replate everything with the proper controls. Unfortunately, we do not have a positive control for erythromycin resistance, but if things die (i.e. negative control works), we should be in business.

Plans for Monday

Lab Work

  • Be sure that the Spc50 plates are working, as the control for Spc50 grew and shouldn't have. My guess (Dan) is that we did not create the stock correctly or used the wrong solvent to dilute it.
  • Finish all the agr PCRs, but especially the RBS.
  • Confirm that amyE insertion works, using the amylase/ery tests.
  • Confirm that with ECE153 inserted, we get detectable fluoresence in a microscope.
  • Confirm that ECE166 transformation works, using plasmid miniprepping.
    • Find the qiagen miniprep kit or adapt the protocol.
  • Confirm that when ECE166 is transformed, we get detectable fluoresence in a microscope.

Book/Computer Work

  • Find good promoters in vectors and order PCR primers to get them out
  • Find out how to assay for/make AIP
    • Find/Contact Jim Ajioka

Longer Term Plans

  • Ligate the RBS into a BB vector so we can build a RBS/GFP construct.
  • Put the RBS/GFP construct into ECE166 and ECE 153.
  • Compare the 'out of the box' ECE153 with ECE153 with RBS/GFP biobrick inserts
  • Compare the 'out of the box' ECE166 with ECE166 with RBS/GFP biobrick inserts




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