"
Team:TU Delft/27 juli 2010 content
From 2010.igem.org
(Difference between revisions)
(New page: Digestion Some BioBricks are in production. Digestion reactions: {| style="color:black; background-color:white;" cellpadding="5"...) |
|||
| Line 53: | Line 53: | ||
loaded 4 μL of marker and 10 μL sample + 2 μL loadingbuffer | loaded 4 μL of marker and 10 μL sample + 2 μL loadingbuffer | ||
| - | [[Image:N 20100727 DIGkopie.jpg]] | + | [[Image:N 20100727 DIGkopie.jpg|300px|thumb|left|Digestion of AlkS, B0015 & E0422]] |
Lane description: | Lane description: | ||
Revision as of 12:59, 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 | XbaI | SpeI | Buffer 2 + BSA (BioLabs) | ’X - AlkS - S’ | |
| 2 | 1.5 μg AlkS | XbaI | SpeI | NCoI | Buffer 2 (BioLabs) | ’X - AlkS - S’ |
| 3 | 1.0 μg B0015 | EcoRI | XbaI | Buffer 2 (BioLabs) | ’E X’- pSB1C3 | |
| 4 | 1.2 μg E0042 | EcoRI | XbaI | Buffer 2 (BioLabs) | ‘E X’- 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
Lane description:
| # | Description | Expected Length (bp) | Status |
| 1 | Marker (5 μL) | ||
| 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
