Thursday 7^th July

Lab work

Bringing DNA closer

By

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

Stock of glycerol

By Mathilde and Laetitia

Transformed cells containing Gblock:

• 1.1 (6clones)
• 1.2 (6clones)
• 2.1(6clones)
• 2.2(6clones)
• 3.1(6clones)
• 3.2(6clones)

For each sample, we made a stock puting 500 of culture and 250 of glycérol (60%)