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===Biobrick characterization=== | ===Biobrick characterization=== | ||
− | ==== ==== | + | ==== Digestion of the plasmid psB1C3 containing the biobrick K1327001==== |
− | ''By '' | + | ''By Mathilde and Alice '' |
+ | |||
+ | For each sample: | ||
+ | 10 microL DNA | ||
+ | 2 L fast digest buffer | ||
+ | 1 L Pst 1 | ||
+ | 6 L H2O | ||
+ | |||
+ | Samples were incubated 5 min at 37°C. | ||
+ | Loading Buffer was added for each solution. Samples were put on an agarose gel for an electrophoresis. | ||
[[File:T--Paris_Saclay--160707_characterization_échelle.jpg|400px|thumb|right|Migration of Cl1(K13) and Cl2 (K13)]] | [[File:T--Paris_Saclay--160707_characterization_échelle.jpg|400px|thumb|right|Migration of Cl1(K13) and Cl2 (K13)]] | ||
+ | Clone 1 didn't show any bands. | ||
+ | Clone 2 showed bands at 2000 kb (corresponding to the size of psB1C3) and 1500 kb (corresponding to the size of biobrick K1327001) | ||
+ | |||
+ | We concluded that clone 2 contain the right insert. | ||
+ | |||
+ | |||
+ | Then, plasmidic DNA of the remaining Clone 1 and 2 (from the extraction of 06/07/16) were re-concentrated using the following protocol: | ||
+ | -Add 2 volumes of cold ethanol (100%) | ||
+ | -Add the quivalent of1/10 of the remaining DNA of solution III | ||
+ | -Put it 30 min at -20°C | ||
+ | -Centrifuge 4 min at1700 rpm | ||
+ | -Remove the supernatant | ||
+ | -Add 1 mL of ethanol at 70% and inverse | ||
+ | -centrifuge 4 min at 1300 rpm | ||
+ | -Let it dry 1 h | ||
+ | -Dilute with 10 L of sterilized water. | ||
+ | -Put it at -20°C | ||
===Visualization=== | ===Visualization=== |
Revision as of 14:40, 27 July 2016