Team:Groningen/Template/MODULE/Notebook/vector/week1

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Instead of <i>mrfp</i>, <i>amil-cp</i> and <i>cp29-rbs-sfgfp</i> were primed with the same primers used for <i>mrfp</i> for pIL253 and pNZ8048g
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This was run on a gel and yielded the right results
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The <i>mrfp</i>, <i>amil-cp</i> and <i>cp29-rbs-sfgfp</i> inserts for pIL253 were ligated to pIL253 and transformed to <i>L. lactis</i> NZ9000 and the <i>amil-cp</i> and <i>cp29-rbs-sfgfp</i> inserts for pNZ8048g were also ligated to pNZ8048g and transformed <i>L. lactis</i> NZ9000
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The transformants of pIL253 were plated on M17 agar with erythromycin and the transformants of pNZ8048g were on M17 agar with chloramphenicol
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This transformation did not yield any colonies
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Revision as of 20:07, 17 October 2014

September 29 - October 5
 
goal: Isolate mrfp insert out of pSB1C3 and ligate in pIL253 and pNZ8048g to make biobrick compatible vectors for L. lactis
 
mrfp was primed out of pSB1C3 by primers designed for pIL253 and pNZ8048g
 
The reactions were loaded on a gel and this yielded positive results for the constructs for both pIL253 and pNZ8048g
 
The backbones pIL253 and pNZ8048g are digested so that the mrfp constructs will fit in them
 
Also the mrfp inserts are digested with the same or complementary enzymes
 
The backbones and mrfp inserts are checked on a gel
 
The gel did not yield good results for the pNZ mrfp
 
Instead of mrfp, amil-cp and cp29-rbs-sfgfp were primed with the same primers used for mrfp for pIL253 and pNZ8048g
 
This was run on a gel and yielded the right results
 
The mrfp, amil-cp and cp29-rbs-sfgfp inserts for pIL253 were ligated to pIL253 and transformed to L. lactis NZ9000 and the amil-cp and cp29-rbs-sfgfp inserts for pNZ8048g were also ligated to pNZ8048g and transformed L. lactis NZ9000
 
The transformants of pIL253 were plated on M17 agar with erythromycin and the transformants of pNZ8048g were on M17 agar with chloramphenicol
 
This transformation did not yield any colonies
 
 
 
 

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