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Team:TU Delft/23 July 2010 content
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| - | With a number of [https://2010.igem.org/Team:TU_Delft#/blog?blog=22_July_2010 yesterday's] colonies a digestion was done to check if they contain the desired insert. The table below describes the digestion procedure, as well as the gel lane description. | + | With a number of [https://2010.igem.org/Team:TU_Delft#/blog?blog=22_July_2010 yesterday's] colonies a digestion was done to check if they contain the desired insert. The table below describes the [https://2010.igem.org/Team:TU_Delft/protocols/restriction_enzyme_digestion digestion] procedure, as well as the gel lane description. (Digestion was done in 10μl end-volume) |
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Revision as of 12:15, 26 July 2010
Alkane Degradation
With a number of yesterday's colonies a digestion was done to check if they contain the desired insert. The table below describes the digestion procedure, as well as the gel lane description. (Digestion was done in 10μl end-volume)
| # | Contents | Enzyme 1 | Enzyme 2 | Buffer | BSA | Expected size(s) (bp) |
| 1 | Smartladder (5μl) | n/a | ||||
| 2 | 007T (2) | 0.5 μl EcoRI | 0.5 μl PstI | 3 (Biolabs) | no | 1341, 2455 |
| 3 | 008T (4) | 0.5 μl EcoRI | 0.5 μl PstI | 3 (Biolabs) | no | 276, 2455 |
| 4 | 009T (6) | 0.5 μl EcoRI | 0.5 μl PstI | 3 (Biolabs) | no | 285, 2455 |
| 5 | 009T (7) | 0.5 μl EcoRI | 0.5 μl PstI | 3 (Biolabs) | no | 285, 2455 |
| 6 | 010T (9) | 0.5 μl EcoRI | 0.5 μl PstI | 3 (Biolabs) | no | 1382, 2455 |
| 7 | 017T (14) | 0.5 μl EcoRI | 0.5 μl PstI | 3 (Biolabs) | no | 855, 2455 |
| 8 | Empty | |||||
| 9 | 007T (2) | 1 μl PstI | 1 μl PvuII | H (Boehringer) | no | 2529,983,251 |
| 10 | 008T (4) | 1 μl AatII | 0.5 μl KpnI | A (Boehringer) | 1 μl | 271, 2427 |
| 11 | 009T (6) | 0.5 μl KpnI | 1 μl NruI | B (Boehringer) | 1 μl | 598, 2109 |
| 12 | 009T (7) | 0.5 μl KpnI | 1 μl NruI | B (Boehringer) | 1 μl | 598, 2109 |
| 13 | 010T (9) | 1 μl EarI | 0.5 μl PstI | 1 (Biolabs) | no | 1075, 2729 |
| 14 | 017T (14) | 0.5 μl AdhI | 0.5 μl SpeI | 4 (Biolabs) | no | 428, 2849 |
| 15 | EZ load (ladder) | n/a |
It seems (from the gel) that BioBrick 007T is the correct one, and the others don't contain the desired insert.
