Team:TU Delft/27 juli 2010 content
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(New page: Digestion Some BioBricks are in production. Digestion reactions: {| style="color:black; background-color:white;" cellpadding="5"...) |
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|- | |- | ||
|1 | |1 | ||
- | |1.5 μg AlkS | + | |1.5 μg AlkS in pSB1C3 |
|XbaI | |XbaI | ||
|SpeI | |SpeI | ||
Line 21: | Line 21: | ||
|- | |- | ||
|2 | |2 | ||
- | |1.5 μg AlkS | + | |1.5 μg AlkS in pSB1C3 |
|XbaI | |XbaI | ||
|SpeI | |SpeI | ||
Line 29: | Line 29: | ||
|- | |- | ||
|3 | |3 | ||
- | |1.0 μg B0015 | + | |1.0 μg B0015 in pSB1AK3 |
|EcoRI | |EcoRI | ||
|XbaI | |XbaI | ||
| | | | ||
|Buffer 2 (BioLabs) | |Buffer 2 (BioLabs) | ||
- | |’E X’- pSB1C3 | + | |’E X’- B0015-pSB1C3 |
|- | |- | ||
|4 | |4 | ||
- | |1.2 μg | + | |1.2 μg E0422 in pS-B1A2 |
|EcoRI | |EcoRI | ||
|XbaI | |XbaI | ||
| | | | ||
|Buffer 2 (BioLabs) | |Buffer 2 (BioLabs) | ||
- | |‘E X’- pSB1C3 | + | |‘E X’- E0422-pSB1C3 |
|- | |- | ||
|} | |} | ||
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Results of the digestion on [[Team:TU_Delft/protocols/agarose_gel|1% agarose gel]] | Results of the digestion on [[Team:TU_Delft/protocols/agarose_gel|1% agarose gel]] | ||
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- | |||
- | |||
- | |||
- | [[Image:N 20100727 DIGkopie.jpg]] | + | [[Image:N 20100727 DIGkopie.jpg|300px|thumb|left|1% Agarose of digestion check, gel runned at 100V for 1 hour, |
+ | loaded 4 μL of marker and 10 μL sample + 2 μL loadingbuffer]] | ||
Lane description: | Lane description: | ||
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|- | |- | ||
|1 | |1 | ||
- | |Marker | + | |Marker |
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Latest revision as of 13:05, 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
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