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Project:Public Biobrick: Difference between revisions
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[[File:Analytical digest public biobrick.jpg]] | [[File:Analytical digest public biobrick.jpg]] | ||
=== Characterisation === | === Characterisation === | ||
Unfortunately, there was no time to characterize (show it work) the biobrick. There is a simple plate assay that could have been used to test the activity of the protein produced. | |||
Protocol given by UCL: | |||
In order to express the gene, the biobrick we have now need to be cut with restriction enzymes to be ligated with a promoter and RBS. It could be characterized by ligating (sticking) onto an expression vector (a plasmid that is commercially available that contains a promoter and ribosome binding site). After transformation do the following: | |||
pick colony from ligation plates into 2ml LB. Incubate @37 until the OD reach approx 0.5. | |||
Perform serial dilutions of half of the liquid culture and plate | |||
Freeze the other half of the culture at -30C overnight. | |||
Next morning, perform the same serial dilutions on the remaining liquid culture and plate. | |||
Count the number of colonies on the plate before freezing and after freezing. | |||
