Team:Groningen/Template/MODULE/Notebook/characterisation/week4

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Fusion of Nis A- terminator and Superfolded GFP-terminator with RBS and purification of the promotor collection (CP1, CP11, CP29, CP30, CP41, CP44)
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Fusion of the NisA with the double terminator and Superfolded GFP with the terminator constructs with the RBS BioBrick (BBa_B0034) and purification of the promotor collection of upsala containing the following promotors: CP1 (BBa_K1033219), CP11 (BBa_K1033220), CP29 (BBa_K1033221), CP30 (BBa_K1033223), CP41 (BBa_K1033224) and CP44 (BBa_K1033225)
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<div class="item">Ligation experiment with PSB2K3 as a backbone and B0034 as 2nd insert</div>
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<div class="item">Both, the NisA with the double terminator and Superfolded GFP with the terminator constructs, were ligated into pSB1K3 together with the RBS BioBrick as a second insert</div>
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<div class="item">Transformation of overnight ligated sample in E.coli</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">Inoculation of the positive clones and plasmid isolation</div>
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<div class="item">Several positive clones were inoculated, and their plasmids were isolated after inoculation</div>
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Revision as of 17:02, 17 October 2014

22 - 28 September
 
Fusion of the NisA with the double terminator and Superfolded GFP with the terminator constructs with the RBS BioBrick (BBa_B0034) and purification of the promotor collection of upsala containing the following promotors: CP1 (BBa_K1033219), CP11 (BBa_K1033220), CP29 (BBa_K1033221), CP30 (BBa_K1033223), CP41 (BBa_K1033224) and CP44 (BBa_K1033225)
 
Both, the NisA with the double terminator and Superfolded GFP with the terminator constructs, were ligated into pSB1K3 together with the RBS BioBrick 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
Several positive clones were inoculated, and their plasmids were isolated after inoculation
 
 
 
 
 

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