Team:Wash U/Calendar-Home/15 July 2009

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(New page: {{WashUback}} == Wednesday July 15, 2009 == -Gel purification from yesterday yielded low DNA concentrations -Started anew today with same digestions but only gel purifying the desti...)
(Wednesday July 15, 2009)
 
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== Wednesday July 15, 2009 ==
 +
-Gel purification from yesterday yielded low DNA concentrations
 +
-Started anew today with same digestions (OmpC, GFP) but only gel purifying the destination plasmids
-
== Wednesday July 15, 2009 ==
+
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
 +
 
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8) 1 kb ladder
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 +
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
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 +
-Made new Kanamycin (10ug/ml) LB plates
 +
 
 +
 
 +
 
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-Gel purification from yesterday yielded low DNA concentrations
 
-
-Started anew today with same digestions but only gel purifying the destination plasmids
 
-
-Team meeting
+
{{WashUbottom}}

Latest revision as of 21:44, 15 July 2009


Wednesday July 15, 2009

-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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