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Project:Public Biobrick: Difference between revisions
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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. | 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: | 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: | 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: | |||
Perform serial dilutions of half of the liquid culture and plate | *Pick colony from ligation plates into 2ml LB. Incubate @37 until the OD reach approx 0.5. | ||
Freeze the other half of the culture at -30C overnight. | *Perform serial dilutions of half of the liquid culture and plate | ||
Next morning, perform the same serial dilutions on the remaining liquid culture and plate. | *Freeze the other half of the culture at -30C overnight. | ||
Count the number of colonies on the plate before freezing and after freezing. | *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. | |||
