Team:Buenos Aires/Results/SynEcoTesting

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=SynEco Testing =
=SynEco Testing =
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=== Coculture of  transformed strains ===
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We did the first experiment to test whether our system works and how two transformed strains grow together.
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We designed an assay to do so; following the growth of these strains:
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{|
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|-
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|[[File:BsAs2012-icono-CFP-202.jpg|200px]]
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|[[File:BsAs2012-icono-YFP-185.jpg|200px]]
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|- align="center"
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|CFP_His
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|YFP_TRPb
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|}
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{|
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|
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{| class="wikitable"
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|+ Transformed cells coculture and controls.
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|! scope="row" align="center" style="background: #7ac5e8"| '''Treatment'''
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|! scope="row" align="center" style="background: #7ac5e8"|'''Strain A'''
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|! scope="row" align="center" style="background: #7ac5e8"| '''Strain B'''
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|-
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|1
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|YFP_TRPb_TRPZipper2
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|CFP_HIS_PolyHa
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|-
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|2
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|YFP_TRPb_TRPZipper2
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|CFP_HIS
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|-
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|3
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|YFP_TRPb_TRPZipper2
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|! scope="row" style="background: #CCCCCC"|
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|-
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|4
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|YFP_TRPb
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|CFP_HIS_PolyHa
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|-
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|5
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|! scope="row" style="background: #CCCCCC"|
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|CFP_HIS_PolyHa
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|-
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|6
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|YFP_TRPb
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|CFP_HIS
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|-
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|7
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|! scope="row" style="background: #CCCCCC"|
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|CFP_HIS
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|-
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|8
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|YFP_TRPb
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|! scope="row" style="background: #CCCCCC"|
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|}
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|}
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==== Protocol ====
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{|
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|
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# Starters of strains were grown over night at 30°C, according the scheme showed in the table
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# The next day cultures were sonicated briefly in low power, and OD was measured in order to check they were in exponential phase.
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# Cells were centrifugated and then washed with medium –HT.
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# We set the culture of strains at OD: 0.02 in 5 ml of medium –HT with 3 replica, according to Table 1.
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# At 0, 1, 2, 3, 4, 5, 6, 7, 8 and 22 hours we took samples of 20 µl.
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# Samples were placed in a 384 wells plate, with 20 µl of cyclohexamide 2x (final concentration 1x) in each of the wells.
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# We used epifluorescence microscope in order to determinate the strain proportion of each coculture. The density of the culture was calculated based on the cell density at in each wells.
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|
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{|class="wikitable"
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|! scope="row" align="center" style="background: #7ac5e8"|'''Strain'''
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|! scope="row" align="center" style="background: #7ac5e8"|'''Media'''
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|-
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|YFP_TRPb_TRPZipper2
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|'''-T'''
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|-
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|YFP_TRPb
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|'''-T'''
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|-
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|CFP_HIS_PolyHa
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|'''-H'''
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|-
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|CFP_HIS
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|'''-H'''
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|}
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|}
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{| class="wikitable" style="width:50%"
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| align="center" | [[File:Bsas2012-Wells.png‎|500px]]
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|-
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| 384 wells plate to be used for epifluorescence microscope.
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|}
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==== Results ====

Revision as of 03:10, 27 October 2012

Contents

SynEco Testing

Coculture of transformed strains

We did the first experiment to test whether our system works and how two transformed strains grow together.

We designed an assay to do so; following the growth of these strains:

BsAs2012-icono-CFP-202.jpg BsAs2012-icono-YFP-185.jpg
CFP_His YFP_TRPb


Transformed cells coculture and controls.
Treatment Strain A Strain B
1 YFP_TRPb_TRPZipper2 CFP_HIS_PolyHa
2 YFP_TRPb_TRPZipper2 CFP_HIS
3 YFP_TRPb_TRPZipper2
4 YFP_TRPb CFP_HIS_PolyHa
5 CFP_HIS_PolyHa
6 YFP_TRPb CFP_HIS
7 CFP_HIS
8 YFP_TRPb

Protocol

  1. Starters of strains were grown over night at 30°C, according the scheme showed in the table
  2. The next day cultures were sonicated briefly in low power, and OD was measured in order to check they were in exponential phase.
  3. Cells were centrifugated and then washed with medium –HT.
  4. We set the culture of strains at OD: 0.02 in 5 ml of medium –HT with 3 replica, according to Table 1.
  5. At 0, 1, 2, 3, 4, 5, 6, 7, 8 and 22 hours we took samples of 20 µl.
  6. Samples were placed in a 384 wells plate, with 20 µl of cyclohexamide 2x (final concentration 1x) in each of the wells.
  7. We used epifluorescence microscope in order to determinate the strain proportion of each coculture. The density of the culture was calculated based on the cell density at in each wells.
Strain Media
YFP_TRPb_TRPZipper2 -T
YFP_TRPb -T
CFP_HIS_PolyHa -H
CFP_HIS -H
Bsas2012-Wells.png
384 wells plate to be used for epifluorescence microscope.


Results

Retrieved from "http://2012.igem.org/Team:Buenos_Aires/Results/SynEcoTesting"