Team:Paris-Saclay/Project/Notebook/Week 8

From 2012.igem.org

(Difference between revisions)

Revision as of 07:12, 26 September 2012

GEMOTE

		Tweet

23rd July

Electrophoresis of the post Gibson assembly PCR on a 1% Agarose gel. We are expecting a band at 4800bp for B, and a band at 1200bp for α and β.
Checking of the purity of BBa_K098995, BBa_K274100 and pSB1A2, all already digested by DPNI

24th July

Purification by column of BBa_K098995, BBa_K274100 and pSB1A2.
Estimation of the concentration of BBa_K098995, BBa_K274100 and pSB1A2 by Nanovue.

Visualization of the purification of BBa_K098995, BBa_K274100 and pSB1A2 by electrophoresis on a 1% Agarose gel. We are expecting a band at 935 bp for BBa_K098995, 3385 pb for BBa_K274100 and 2079 bp for pSB1A2.

25th July

Amplification by PCR of BBa_K098995, BBa_K274100 and pSB1A2. Visualization by electrophoresis on a 1% Agarose gel. We are expecting a band at 935 bp for BBa_K098995, 3385 pb for BBa_K274100 and 2079 bp for pSB1A2.

PCR program used:

Treatment by DPNI on BBa_K098995, BBa_K274100 and pSB1A2 in order to eliminate the plasmid matrix.

26th July

Purification of BBa_K098995, BBa_K274100 and pSB1A2 with the gel extraction PCR clean up kit to prepare a Gibson assembly.
Nanovue to measure the concentration of our 3 samples: BBa_K098995, BBa_K274100 and pSB1A2.

27th July

Gibson assembly for the B construction (BBa_K115017 + 880bp end of BBa_K098995 + BBa_K274100 + BBa_J61048 + pSB1A2). A control is done with just the plasmid treated by DPNI.
Transformation of competent cell DH5α with the B construction. Spreading out on LB+Agar+Ampicilline, and the Petri dishes are placed at 37°C.

		Tweet

@@ Line 77: / Line 77: @@
 [[File:Week8-5.jpg|400px]]
 PCR program used:
-[[File:Week8-6.jpg|600px]]
+[[File:Week8-6.jpg|650px]]
 *Treatment by DPNI on BBa_K098995, BBa_K274100 and pSB1A2 in order to eliminate the plasmid matrix.