Team:Groningen/Template/MODULE/Notebook/characterisation/week3

From 2014.igem.org

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Fusion of Nis A and Superfolded GFP with terminator
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Fusion of the NisA BioBrick (BBa_K1365000) and the Superfolded GFP BioBrick (BBa_K1365020) with a double terminator (BBa_B0015)
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<div class="item">Single and double digestion of Nis A and Superfolded GFP to confirm the size of the plasmid</div>
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<div class="item">The size of the plasmids were confirmed by single and double digestion of the plasmids (pSB1C3) with the NisA and Superfolded GFP BioBricks to confirm the sizes of the plasmids</div>
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<div class="item">Ligation experiment with PSB1A3 as a backbone and B0015 as 2nd insert</div>
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<div class="item">Both, the NisA and the Superfolded GFP BioBrick, were ligated into pSB1A3 together with the double terminator as a second insert</div>
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<div class="item">Transformation of overnight ligated sample in <i>E. coli</i></div>
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<div class="item">The ligation was performed overnight, after which the ligated plasmids were transformed to two different <i>E. coli</i> DH5α strains</div>
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<div class="item">Size of the construct was determined with the help of colony PCR</div>
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<div class="item">The size of the construct was determined by colony PCR</div>
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<div class="item">Double digestion, purification of the construct</div>
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<div class="item">A double digestion was performed, and the constructs were further purified</div>
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Revision as of 16:40, 17 October 2014

15 - 21 September
 
Fusion of the NisA BioBrick (BBa_K1365000) and the Superfolded GFP BioBrick (BBa_K1365020) with a double terminator (BBa_B0015)
 
The size of the plasmids were confirmed by single and double digestion of the plasmids (pSB1C3) with the NisA and Superfolded GFP BioBricks to confirm the sizes of the plasmids
Both, the NisA and the Superfolded GFP BioBrick, were ligated into pSB1A3 together with the double terminator as a second insert
The ligation was performed overnight, after which the ligated plasmids were transformed to two different E. coli DH5α strains
The size of the construct was determined by colony PCR
A double digestion was performed, and the constructs were further purified
 
 
 
 
 

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