Team:UNITN-Trento/Notebook/Labposts/08/24

From 2013.igem.org

{ "date" : "2013-08-10", "author" : "viola-thomas", "title" : "cloning of s04617+EFE+B0015 and AraCpBAD+EFE+B0015 in psb1c3", "content" : "Starting from the overnight digestion of yesterday of the EFE PCR with XbaI and PstI (as insertion)and the parts s04617 and AraCpBAD with SpeI and PstI (as destination plasmid)today we treated the isert with DpnI and the plasmids with SAP. We purified the digestions and the final quantification were :

Quatification results
partinitial amount digestedquantification after purification
EFEall PCR19.5 ng/µl
AraCpBAD3µg38.3 ng/µl
s046173µg13µg
Then we ligate following this protocol. With the part s04617 (pFixK-cI-pLambda) we could do only the 1:2 ligation and the control because of the very small amount of destination vector. Then we transformed the Neb5α cells following the classical transformation protocol.", "tags" : "pFixK-cI-pLambda-EFE-AraCpBAD" }