IGEM:IMPERIAL/2009/Assays Protocols: Difference between revisions
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=Current Wet Lab Plan= | |||
[[IGEM:IMPERIAL/2009/Assays_Protocols/WetLab_Plan | Step By Step Wet-Lab Plan]] | |||
= Assays = | = Assays = | ||
==Functional Assays== | |||
<b>Capsule: Is it able to withstand Stomach pH and protease conditions? </b> | |||
We must test the functionality of the complete capsule, to see if it can withstand the conditions present in the stomach. | |||
This can be done experimentally by recreating the stomach conditions in the lab, and seeing the effects on the bacteria. This could include quantifying the number of bacteria broken down. | |||
<b>Trehalose Functionality: Does it help protect against dessication? </b> | |||
As well as testing for the presence of trehalose, it may be useful to test to see if it performs its required function within our cells ie to protect against dessicating conditions. | |||
* As trehalose is naturally present in E.coli, is it ok to assume that its presence will protect against dessication? | |||
<br> | |||
==Shopping Lists!== | |||
[[IGEM:IMPERIAL/2009/Assays_Protocols/Shopping | Shopping]] | |||
==Module 1== | ==Module 1== | ||
=== Cellulase === | === Cellulase === | ||
Cellulase Assay from Sigma Aldrich <br> | Cellulase Assay from Sigma Aldrich <br> | ||
[[Media:Cellulase Assay.pdf | Cellulase Assay (Sigma)]] | [[Media:Cellulase Assay.pdf | Cellulase Assay (Sigma)]] <b>Quantitative</b> | ||
<br><br> | <br><br> | ||
Feedback - In vitro or vivo? Do we need to purify enzyme for assay? presumably so, as requires measurement of glucose concentation, therefore not good to use cell extract </i> | |||
<i>Feedback - In vitro or vivo? Do we need to purify enzyme for assay? presumably so, as requires measurement of glucose concentation, therefore not good to use cell extract </i> | |||
<br> | <br> | ||
=== Phenylalanine Hydroxylase (PAH)=== | === Phenylalanine Hydroxylase (PAH)=== | ||
Phenylalanine Hydroxylase Assay from Sigma Aldrich : <br> | Phenylalanine Hydroxylase Assay from Sigma Aldrich : <br> | ||
[[Media:PAH Assay.pdf | PAH Assay]] | [[Media:PAH Assay.pdf | PAH Assay]] <b>Quantitative</b> | ||
==Module 2== | ==Module 2== | ||
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[[IGEM:IMPERIAL/2009/Assays_Protocols/Trehalose | Trehalose Assay]] | [[IGEM:IMPERIAL/2009/Assays_Protocols/Trehalose | Trehalose Assay]] | ||
<br><br> | <br><br> | ||
==Module 3== | |||
[[IGEM:IMPERIAL/2009/Assays_Protocols/M3_heatinduction |Heat induction Assays]] | |||
[[IGEM:IMPERIAL/2009/Assays_Protocols/M3_Assays |Cell Death Assays]] | |||
=Cloning Strategies= | =Cloning Strategies= | ||
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[[IGEM:IMPERIAL/2009/Assays_Protocols/Cloning_strategy_M2 | Cloning Strategy for Module 2]] | [[IGEM:IMPERIAL/2009/Assays_Protocols/Cloning_strategy_M2 | Cloning Strategy for Module 2]] | ||
==Testing Constructs== | |||
[[IGEM:IMPERIAL/2009/Assays_Protocols/Testing_Constructs| Testing Constructs for all modules]] | |||
Latest revision as of 06:46, 7 August 2009
Current Wet Lab Plan
Assays
Functional Assays
Capsule: Is it able to withstand Stomach pH and protease conditions? We must test the functionality of the complete capsule, to see if it can withstand the conditions present in the stomach. This can be done experimentally by recreating the stomach conditions in the lab, and seeing the effects on the bacteria. This could include quantifying the number of bacteria broken down.
Trehalose Functionality: Does it help protect against dessication? As well as testing for the presence of trehalose, it may be useful to test to see if it performs its required function within our cells ie to protect against dessicating conditions.
- As trehalose is naturally present in E.coli, is it ok to assume that its presence will protect against dessication?
Shopping Lists!
Module 1
Cellulase
Cellulase Assay from Sigma Aldrich
Cellulase Assay (Sigma) Quantitative
Feedback - In vitro or vivo? Do we need to purify enzyme for assay? presumably so, as requires measurement of glucose concentation, therefore not good to use cell extract
Phenylalanine Hydroxylase (PAH)
Phenylalanine Hydroxylase Assay from Sigma Aldrich :
PAH Assay Quantitative
Module 2
Module 3
Cloning Strategies
Cloning Strategy for Module 1 and Timer
