Team:HKU-HKBU/Motor Preliminary Trials

From 2009.igem.org

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(Binding performance test using a membrane)
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==Binding performance test using a membrane==
==Binding performance test using a membrane==
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We used Immobilon-P transfer membrane to evaluate the performance of streptavidin-biotin binding of bacteria onto a surface. Before cutting apart the membrane into small pieces, pre-activation should be applied to it first  (see protocols [[Team:HKU-HKBU/Protocols#Membrane_Biotinylation | membrane biotinylation]]). After the pre-activation, the membrane was first sheared into strings manually by using scissors, with the width and thickness being approximately 100μm. We then used Leica-crytomicrotome to cut the “threads” into even smaller fragments, with the length of which being 60μm. A binding performance testing device demension was aproximately 100μm×60μm×100μm. Since the surface of membrane had already been activated, the polar-expression bacteria could bind onto such biotin-coated motors.  
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We used Immobilon-P transfer membrane to evaluate the performance of streptavidin-biotin binding of bacteria onto a surface. Before cutting apart the membrane into small pieces, pre-activation was applied to it first  (see protocol [[Team:HKU-HKBU/Protocols#Membrane_Biotinylation | membrane biotinylation]]). After the pre-activation, the membrane was first sheared into strings manually by scissors, with the width and thickness being approximately 100μm. We then used Leica-crytomicrotome to cut the “threads” into even smaller fragments, with the length of which being 60μm. The dimension of the binding performance testing device is aproximately 100μm×60μm×100μm. Since the surface of membrane had already been activated, the polar-expression bacteria could bind onto such biotin-coated motors.  
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[[Image:HKU-HKBU_motor_results_1.png| left | thumb |200px|Step1. The fragmentation of the Immobolin-P membrane with the help of a mini-homogenizer]]
[[Image:HKU-HKBU_motor_results_1.png| left | thumb |200px|Step1. The fragmentation of the Immobolin-P membrane with the help of a mini-homogenizer]]

Revision as of 11:24, 21 October 2009

Binding performance test using a membrane

We used Immobilon-P transfer membrane to evaluate the performance of streptavidin-biotin binding of bacteria onto a surface. Before cutting apart the membrane into small pieces, pre-activation was applied to it first (see protocol membrane biotinylation). After the pre-activation, the membrane was first sheared into strings manually by scissors, with the width and thickness being approximately 100μm. We then used Leica-crytomicrotome to cut the “threads” into even smaller fragments, with the length of which being 60μm. The dimension of the binding performance testing device is aproximately 100μm×60μm×100μm. Since the surface of membrane had already been activated, the polar-expression bacteria could bind onto such biotin-coated motors.

Step1. The fragmentation of the Immobolin-P membrane with the help of a mini-homogenizer
Step2.The fragmentation of the Immobolin-P membrane with a generated shearing force by moistening and followed by vortexing

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