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

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