Thursday 7^th July

Lab work

Bringing DNA closer

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Migration of NM, TD, ST1, ST2, SP, TD, pTet

Biobrick characterization

Digestion of the plasmid psB1C3 containing the biobrick K1327001

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.

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

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