Difference between revisions of "Team:Austin UTexas/HP/Gold Integrated"

Revision as of 03:53, 16 October 2018

Integrated Human Practices

Optimization of Golden Gate Assembly

In order to increase the transformation efficiency of Golden Gate Assembly reactions we made standardized bridging sequences that reduced the number of parts in a reaction, increasing the likelihood of a successful assembly. Using this method, we were able to make many assemblies expressing different antibiotic resistance quickly.

**Fig 1.** Schematic of teaction to make a 1-5 Bridge. The 1-5 Bridge reduces the number of parts in a BsaI Golden Gate assembly reaction to make a full plasmid that contains all part types.

Transformation of Assembly Plasmids into Mu Free Donors

Dr.Brian Redna of Gingko Bioworks emphasized that our kit would be limited by the ability of the plasmids to be inserted into the bacteria of interest as many can not be transformed with standard protocols. Therefore, we transformed the assembly plasmids into a strain of E. coli that can act as a plasmid donor in conjugations.

See Demonstrate Page

**Figure 1:** These are the results of Rice's one tube electroporation with *E. coli*.

@@ Line 29: / Line 29: @@
 <h3> Optimization of Golden Gate Assembly </h3>
 <p><b>In order to increase the transformation efficiency of Golden Gate Assembly reactions we made standardized bridging sequences that reduced the number of parts in a reaction, increasing the likelihood of a successful assembly. Using this method, we were able to make many assemblies expressing different antibiotic resistance quickly.</b></p>
+<figure><img class = "center", src="https://static.igem.org/mediawiki/2018/8/84/T--Austin_UTexas--1-5Bridge.JPG">
+<figcaption><b> Fig 1. </b> Schematic of teaction to make a 1-5 Bridge. The 1-5 Bridge reduces the number of parts in a BsaI Golden Gate assembly reaction to make a full plasmid that contains all part types.</figcaption>
+</figure>
 <h3>Transformation of Assembly Plasmids into Mu Free Donors</h3>