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Team:University of Lethbridge/Notebook/GeneralLabOctober
From 2008.igem.org
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October 1, 2008
Roxanne
-Inactivated the Enzymes in the morning
-Ran a gel of the tested pLacI, along with the previously cut RFP and TetR in the afternoon on 1% Agarose Gel in TAE.
-the iGEM pLacI had a faint band, RFP looks good (half of it cut), TetR looks like it either didn't cut at all, or only cut once. Only used 2 uL for each sample, will try running with more.
October 4, 2008
Roxanne
-Reran the pLacI tests, RFP and TetR on a 1% Agarose Gel in TAE.
-Repicked pLacI x2, RFP and TetR colonies into LB+amp media since I've been having trouble with ligations. Brent suggested using lots of DNA in the ligation (<100 ng) to make ure that the ligation does in fact work this time.
October 5, 2008
Roxanne
-plasmid prepped the pLacI x2, RFP and TetR subcultures. Lost one of the pLacI cultures (cells wouldn't lyse). Had some left over pLacI - 2 culture, attempted to plasmid prep those cells.
-Ran a gel of the plasmid preps.
-Restriction Digest the pLacI - 1, RFP and TetR plasmids obtained today, as well as recutting the TetR plasmid from last time, since it appears as though it didn't cut at all, or only cut once. I will be using the iGEM enzymes. Left to cut overnight at 37.0C
-picked some pLacI colonies from a different plate. Maybe I'll have better luck there. Incubate in LB+amp media at 37.0C for 15 hours.
