Team:TU Delft/27 juli 2010 content

From 2010.igem.org

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|1.5 μg AlkS
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|1.5 μg AlkS in pSB1C3
|XbaI
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|SpeI
|SpeI
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|1.5 μg AlkS  
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|1.5 μg AlkS in pSB1C3
|XbaI
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|SpeI
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|1.0 μg B0015
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|1.0 μg B0015 in pSB1AK3
|EcoRI
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|XbaI
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|Buffer 2 (BioLabs)
|Buffer 2 (BioLabs)
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|’E X’- pSB1C3  
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|’E X’- B0015-pSB1C3  
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|1.2 μg E0042
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|1.2 μg E0422 in pS-B1A2
|EcoRI
|EcoRI
|XbaI
|XbaI
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|Buffer 2 (BioLabs)
|Buffer 2 (BioLabs)
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|‘E X’- pSB1C3
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|‘E X’- E0422-pSB1C3
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|Marker (5 μL)   
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|Marker
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Revision as of 13:03, 28 July 2010

Digestion

Some BioBricks are in production. Digestion reactions:

# Sample Enzyme 1 Enzyme 2 Enzyme 3 Buffer Needed fragment
1 1.5 μg AlkS in pSB1C3 XbaI SpeI Buffer 2 + BSA (BioLabs) ’X - AlkS - S’
2 1.5 μg AlkS in pSB1C3 XbaI SpeI NCoI Buffer 2 (BioLabs) ’X - AlkS - S’
3 1.0 μg B0015 in pSB1AK3 EcoRI XbaI Buffer 2 (BioLabs) ’E X’- B0015-pSB1C3
4 1.2 μg E0422 in pS-B1A2 EcoRI XbaI Buffer 2 (BioLabs) ‘E X’- E0422-pSB1C3


Results of the digestion on 1% agarose gel Title figure: 1% Agarose of digestion check gel runned at 100V for 1 hour loaded 4 μL of marker and 10 μL sample + 2 μL loadingbuffer

Digestion of AlkS, B0015 & E0422

Lane description:

# Description Expected Length (bp) Status
1 Marker
2 Undigested B0015 in pSB1AK3 3318 ✓
3 Digested B0015 in pSB1AK3 + EcoRI + XBaI 3303, 15 ✓
4 Undigested E0422 in pSB1A2 2996 ✓
5 Digested E0422 in pSB1A2 + EcoRI + XBaI 2981, 15 ✓
6 Undigested AlkS in pSB1C3 4999 ✓
7 Digested AlkS in pSB1C3 + EcoRI + SpeI + NCoI 2673, 1249, 1077 ✓
8 Digested AlkS in pSB1C3 + EcoRI + SpeI 2673, 2326 ✓

Conclusion: All the plasmids were digested as expected

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