Team:TU Delft/13 July 2010 content

From 2010.igem.org

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|1
|1
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|Biorad EZ marker
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|Biorad EZ marker (5 μL)
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|2
|2
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|Undigested pSB1T3
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|Undigested pSB1T3 (10 μL + 2 μL loadingbuffer)
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|3
|3
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|Digested pSB1T3
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|Digested pSB1T3 (10 μL + 2 μL loadingbuffer)
|2500
|2500
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|4
|4
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|Undigested AlnA
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|Undigested AlnA (10 μL + 2 μL loadingbuffer)
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|5
|5
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|Digested AlnA
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|Digested AlnA (10 μL + 2 μL loadingbuffer)
|1107
|1107
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|6
|6
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|Undigested OprG
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|Undigested OprG (10 μL + 2 μL loadingbuffer)
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|7
|7
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|Digested OprG
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|Digested OprG (10 μL + 2 μL loadingbuffer)
|744
|744
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|8
|8
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|Undigested B0015
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|Undigested B0015 (10 μL + 2 μL loadingbuffer)
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|9
|9
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|Digested B0015
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|Digested B0015 (10 μL + 2 μL loadingbuffer)
|130
|130
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|10
|10
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|Undigested R0011 '''!'''
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|Undigested R0011 (10 μL + 2 μL loadingbuffer) '''!'''
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|Digested R0011  
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|Digested R0011 (10 μL + 2 μL loadingbuffer)
|55
|55
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|12
|12
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|Undigested B0032
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|Undigested B0032 (10 μL + 2 μL loadingbuffer)
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|13
|13
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|Digested B0032
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|Digested B0032 (10 μL + 2 μL loadingbuffer)
|13
|13
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|14
|14
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|SmartLadder
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|SmartLadder (5 μL)
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Revision as of 20:17, 18 July 2010

Lab work

Emulsifier

Today the digestion products from yesterday were run on gel to see whether the plasmids were cut in the right way.

1% Agarose gel runned at 100V for 1h
EuroGentec SmartLadder

Lane description

# Description Expected Length (bp)
1 Biorad EZ marker (5 μL)
2 Undigested pSB1T3 (10 μL + 2 μL loadingbuffer)
3 Digested pSB1T3 (10 μL + 2 μL loadingbuffer) 2500
4 Undigested AlnA (10 μL + 2 μL loadingbuffer)
5 Digested AlnA (10 μL + 2 μL loadingbuffer) 1107
6 Undigested OprG (10 μL + 2 μL loadingbuffer)
7 Digested OprG (10 μL + 2 μL loadingbuffer) 744
8 Undigested B0015 (10 μL + 2 μL loadingbuffer)
9 Digested B0015 (10 μL + 2 μL loadingbuffer) 130
10 Undigested R0011 (10 μL + 2 μL loadingbuffer) !
11 Digested R0011 (10 μL + 2 μL loadingbuffer) 55
12 Undigested B0032 (10 μL + 2 μL loadingbuffer)
13 Digested B0032 (10 μL + 2 μL loadingbuffer) 13
14 SmartLadder (5 μL)

! Sample not fully loaded on gel

The band position of the digestion products of AlnA and OprG correspond to their length. The other fragments (R0011, B0032 and B0015) probably run of the gel.

The ligation products that were incubated over night were transformed to Top10 competent cells according to the protocol.

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