"
29/07/13
From 2013.igem.org
(Difference between revisions)
| Line 3: | Line 3: | ||
*Three different reactions | *Three different reactions | ||
*Master Mix (changes were made from the protocol) | *Master Mix (changes were made from the protocol) | ||
| - | **5ul NEB Buffer | + | **5ul NEB Buffer 3.1 |
**0.5ul of EcoRI | **0.5ul of EcoRI | ||
**0.5ul of PstI | **0.5ul of PstI | ||
| Line 10: | Line 10: | ||
**4ul of linearized backbone | **4ul of linearized backbone | ||
**4ul of enzyme master mix | **4ul of enzyme master mix | ||
| + | *digest at 37C for 30min | ||
| + | *heat kill at 80C for 20min | ||
| + | #Digestion of the limonene biobrick plasmid backbone (BBa_K118025) | ||
| + | *Using clone 1.1 purified plasmid DNA | ||
| + | *For the reaction adding: | ||
| + | **5ul NEB Buffer 3.1 | ||
| + | **2ul of EcoRI | ||
| + | **2ul of PstI | ||
| + | **2.5ul of dH2O | ||
| + | **38.5ul of DNA | ||
*digest at 37C for 30min | *digest at 37C for 30min | ||
*heat kill at 80C for 20min | *heat kill at 80C for 20min | ||
Revision as of 14:10, 29 July 2013
- Ran the gel from the 26/07/2013 again for 45min
- Digestion of plasmid backbone (pSB1C3)
- Three different reactions
- Master Mix (changes were made from the protocol)
- 5ul NEB Buffer 3.1
- 0.5ul of EcoRI
- 0.5ul of PstI
- 19ul of dH2O
- For each reaction add:
- 4ul of linearized backbone
- 4ul of enzyme master mix
- digest at 37C for 30min
- heat kill at 80C for 20min
- Digestion of the limonene biobrick plasmid backbone (BBa_K118025)
- Using clone 1.1 purified plasmid DNA
- For the reaction adding:
- 5ul NEB Buffer 3.1
- 2ul of EcoRI
- 2ul of PstI
- 2.5ul of dH2O
- 38.5ul of DNA
- digest at 37C for 30min
- heat kill at 80C for 20min
