Difference between revisions of "Part:BBa K1111004:Experience"

Revision as of 16:48, 2 October 2013

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_K1111004

We wanted to test if the promoter that we cloned into the plasmid really can be induced upon external acidification. In order to do that, we transformed bacteria with it, incubated them at different pH and then measured their GFP expression using a plate reader.

Gibson Assembly and Primers

We used Gibson Assembly to insert the Cad promoter into the pSB1C3 Backbone. We used the biobrick BBa_I746908 where we remove the Arac promoter but left the superfolded GFP. This way, the latter was under control of our Cad promoter.

File:1.2 History

Figure 1: History of the Gibson Assembly including Primers

Sequencing

The sequencing result showed that there were no mutations in the insert

Figure 1: Sequencing result of the Cad promoter using one foreward Primer that binds directly to the beginning of the promoter

OD measurements

We transformed cell with our plasmid and then measured their OD in different media with different pHs.

File:CADOD.jpg

Figure 1: Growth Curve of transformed DH5-alpha cells in different Media

User Reviews

UNIQc41fff94394e0152-partinfo-00000000-QINU UNIQc41fff94394e0152-partinfo-00000001-QINU

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-===Primers===
+===Gibson Assembly and Primers===
+We used Gibson Assembly to insert the Cad promoter into the pSB1C3 Backbone. We used the biobrick BBa_I746908 where we remove the Arac promoter but left the superfolded GFP. This way, the latter was under control of our Cad promoter.
+[[Image: 1.2_History|thumb|600px|center|Figure 1: History of the Gibson Assembly including Primers]]
 ===Sequencing===