IGEM:IMPERIAL/2009/Assays Protocols

=Current Wet Lab Plan= Step By Step 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?

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Cellulase
Cellulase Assay from Sigma Aldrich [[Media:Cellulase Assay.pdf | 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 : [[Media:PAH Assay.pdf | PAH Assay]]  Quantitative

Module 2
Colanic Acid Assay

Trehalose Assay

Module 3
Heat induction Assays

Cell Death Assays

=Cloning Strategies=

Cloning Strategy for Module 1 and Timer

Cloning Strategy for Module 2

Testing Constructs
Testing Constructs for all modules