Team:Judd UK/Pages/Results
Results
Example
First Culture of Colonies
One of the first tests we performed on our plasmids was to
produce colonies of E. coli which did not produce the chromoprotein. To do
this, we cultured our bacteria on ampicillin agar that had a Fe2+ concentration
of 0.1 M, however we were unable to observe any growth compared to our control
experiment. After further research, we discovered that lethal iron
concentrations for the bacteria were just 1mM<a href="file:///N:/Results%20(1).docx#_ftn1" name="_ftnref1" title="">[1]</a> so
Determining the identity of our constructs
<img src="
">On the right is our attempt to transfer our constructs from the ampicillin resistant to a chloramphenicol resistant plasmid; we generated several fragments during digestion of plasmids 1, 2 and 3 which when ligated with the chloramphenicol resistant plasmid could have produced 2 different possible outcomes. In order to confirm the identity of our constructs, we extracted DNA from colonies using a miniprep kit. However, the identities of construct 2 and construct 3 were likely to be correct as they would appear blue in the absence of iron, (there is no Fe2+ to cause dimerization of the FUR protein and therefore no inhibition of amilCP synthesis in ligation 3 and there is no LacI protein present in our samples which would also inhibit the production of amilCP in ligation 2 therefore colonies that had the correct plasmid would appear blue) and ran a gel electrophoresis on our sample. Using EcoR1 and Pst1 to generate our fragments we expected to generate (insert research here regarding base pairs) and tests 1.1, 1.3, 2.1, 2.2 and 3.1 confirmed the identity of our constructs
<a href="file:///N:/Results%20(1).docx#_ftnref1" name="_ftn1" title="">[1]</a> http://www.bioline.org.br/request?se08030<o:p></o:p>
