Revision as of 00:39, 27 September 2012

GEMOTE Project

Week 11

Purification by PCR clean-up of BBa_K115017 treated with DPNI. To visualize the fragment, an electrophoresis has been made with a 2% Agarose gel. We are expecting a band at the size of 123 bp.
	New PCR of BBa_K115017 to obtain more quantity of the fragment

PCR program used:

Determination of the concentration of plasmid pSB1A2 which has been linearized by HINDIII. We are expecting a band at size (2079+935) ~3kbp
A new PCR of the plasmid pSB1A2 is realized to obtain more quantity of the plasmid. We are expecting a band at size 2079 bp.

PCR program used:

Determination of the concentration of the BBa_K274100 by electrophoresis on a gel at O.8% Agarose. We are expecting a band at size 2100bp

Digestion of the BBa_K274100 by DPNI to degrade the matrix plasmid.
Stocking up cells with glycerol
- PSB1A3Amil CP + Ampicilline
- PSB1C3 Amil GFP + Chloramphinicol

French holiday.

PCR of BBa_K274100 and visualization by 0.8% Agarose gel electrophoresis. We are expecting a band at 3400bp.

PCR program used:

Visualization of the DPNI’s digestion of the plasmid pSB1A2 by 0.8% Agarose gel electrophoresis. . We are expecting a band at size 2079 bp.

Miniprep of the plasmid pSB1A2.
Miniprep of BBa_K274100 and BBa_K115017
Gibson assembly of the B construction
Transformation of DH5α competent cells with the Gibson B construction. The Petri dishes are placed at 37°C.

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 {| width="85%"
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-| style="width: 50%;"|Visualization of the DPNI’s digestion of the plasmid pSB1A2 by 0.8% Agarose gel electrophoresis. . We are expecting a band at the size of 2079 bp.
+| style="width: 50%;"|Visualization of the DPNI’s digestion of the plasmid pSB1A2 by 0.8% Agarose gel electrophoresis. . We are expecting a band at size 2079 bp.
 | style="width: 35%;"| [[File:Week11-10.jpg|right|220px]]
 |}