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Team:Wash U/Calendar-Home/15 July 2009
From 2009.igem.org
-Gel purification from yesterday yielded low DNA concentrations
-Started anew today with same digestions (OmpC, GFP) but only gel purifying the destination plasmids
Gel layout:
1) 100 bp ladder
2) OmpC + vector (2187 bp)
3) OmpC + vector
4) GFP (876 bp)
5) GFP
6) Destination plasmid, pSB1K3 (2206 bp)
7) pSB1K3
8) 1 kb ladder
Gel #2:
1) 1 kb ladder
2) OmpC + vector (2187 bp)
3) same
4) same
5) same
6) GFP (876 bp)
7) same
Note: This second digestion was run using the GFP sample from the 7-1 miniprep and the OmpC sample from the 7-2 miniprep. The 7-2 OmpC DNA was very dilute and required about 74 microliters to obtain 700 ng, so enzyme and buffer volumes were adjusted for a total reaction volume of 100 microliters (requiring four gel lanes).
Ligation reactions:
1: 6 microliters OmpC, 8 microliters GFP, 4 microliters pSB1K3, 2 microliters buffer, 1 microliter enzyme
2: 4 microliters OmpC, 6 microliters GFP, 4 microliters pSB1K3, 2 microliters buffer, 1 microliter enzyme, 3 microliters water
-Team meeting #6
-Made new Kanamycin (10ug/ml) LB plates
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